Methods and compositions for treating amyloid-related diseases

ABSTRACT

Methods, compounds, pharmaceutical compositions and kits are described for treating or preventing amyloid-related disease.

RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationNo. 60/638,636, filed on Dec. 22, 2004. This application is related toU.S. patent application Ser. No. 10/871,514, filed Jun. 18, 2004, whichclaims priority to U.S. patent application Ser. No. 10/871,365 filedJun. 18, 2004, U.S. Provisional Patent Application No. 60/512,047, filedOct. 17, 2003, and U.S. Provisional Patent Application No. 60/480,906,filed Jun. 23, 2003, all entitled Methods and Compositions for TreatingAmyloid-Related Diseases. This application is also related to U.S.Provisional Patent Application 60/638,819, entitled Methods andCompositions for Treating Amyloid-Related Diseases, filed Dec. 22, 2004and U.S. Application Serial No. 11/XXX,XXX, filed concurrently herewith.

This application is also related to U.S. Provisional Patent ApplicationNo. 60/512,017, filed Oct. 17, 2003, U.S. Provisional Patent ApplicationNo. 60/480,918, filed Jun. 23, 2003, and U.S. patent application Ser.No. 10/871,613, filed Jun. 18, 2004, all entitled Methods for TreatingProtein Aggregation Disorders.

This application is related to U.S. Provisional Patent Application No.60/512,116, filed Oct. 17, 2003, U.S. Provisional Patent Application No.60/480,984, filed Jun. 23, 2003, and U.S. application Ser. No.10/871,549, filed Jun. 18, 2004, all entitled PharmaceuticalFormulations of Amyloid-Inhibiting Compounds.

This application is related to U.S. Provisional Patent Application No.60/436,379, filed Dec. 24, 2002, U.S. Provisional Patent Application No.60/482,214, filed Jun. 23, 2003, entitled Combination Therapy for theTreatment of Alzheimer's Disease, U.S. patent application Ser. No.10/746,138, filed Dec. 24, 2003, International Patent Application No.PCT/CA2003/002011, filed Dec. 24, 2003, and U.S. patent application Ser.No. 10/871,537, filed Jun. 18, 2004, entitled Therapeutic Formulationsfor the Treatment of Beta-Amyloid Related Diseases.

This application is related to U.S. Provisional Patent Application No.60/512,135, filed Oct. 17, 2003, U.S. Provisional Patent Application No.60/482,058, filed Jun. 23, 2003, both entitled Synthetic Process forPreparing Compounds for Treating Amyloidosis, and U.S. patentapplication Ser. No. 10/871,543, filed Jun. 18, 2004, entitled ImprovedPharmaceutical Drug Candidates and Method for Preparation Thereof.

This application is related to U.S. Provisional Patent ApplicationSerial No. 60/512,018, filed on Oct. 17, 2003 and U.S. ProvisionalPatent Application Ser. No. 60/480,928, filed on Jun. 23, 2003, and U.S.application Ser. No. 10/871,512, filed Jun. 18, 2004, all entitledMethods and Compositions for Treating Amyloid-andEpileptogenesis-Associated Diseases.

This application is also related to Method for Treating Amyloidosis,U.S. patent application Ser. No. 08/463,548, now U.S. Pat. No.5,972,328.

The entire contents of each of these patent applications and patents arehereby expressly incorporated herein by reference including withoutlimitation the specification, claims, and abstract, as well as anyfigures, tables, or drawings thereof.

BACKGROUND

Amyloidosis refers to a pathological condition characterized by thepresence of amyloid fibrils. Amyloid is a generic term referring to agroup of diverse but specific protein deposits (intracellular orextracellular) which are seen in a number of different diseases. Thoughdiverse in their occurrence, all amyloid deposits have commonmorphologic properties, stain with specific dyes (e.g., Congo red), andhave a characteristic red-green birefringent appearance in polarizedlight after staining. They also share common ultrastructural featuresand common X-ray diffraction and infrared spectra.

Amyloid-related diseases can either be restricted to one organ or spreadto several organs. The first instance is referred to as “localizedamyloidosis” while the second is referred to as “systemic amyloidosis.”

Some amyloid diseases can be idiopathic, but most of these diseasesappear as a complication of a previously existing disorder. For example,primary amyloidosis (AL amyloid) can appear without any other pathologyor can follow plasma cell dyscrasia or multiple myeloma.

Secondary amyloidosis is usually seen associated with chronic infection(such as tuberculosis) or chronic inflammation (such as rheumatoidarthritis). A familial form of secondary amyloidosis is also seen inother types of familial amyloidosis, e.g., Familial Mediterranean Fever(FMF). This familial type of amyloidosis is genetically inherited and isfound in specific population groups. In both primary and secondaryamyloidosis, deposits are found in several organs and are thusconsidered systemic amyloid diseases.

“Localized amyloidoses” are those that tend to involve a single organsystem. Different amyloids are also characterized by the type of proteinpresent in the deposit. For example, neurodegenerative diseases such asscrapie, bovine spongiform encephalitis, Creutzfeldt-Jakob disease, andthe like are characterized by the appearance and accumulation of aprotease-resistant form of a prion protein (referred to as AScr orPrP-27) in the central nervous system. Similarly, Alzheimer's disease,another neurodegenerative disorder, is characterized by neuritic plaquesand neurofibrillary tangles. In this case, the amyloid plaques found inthe parenchyma and the blood vessel is formed by the deposition offibrillar Aβ amyloid protein. Other diseases such as adult-onsetdiabetes (type II diabetes) are characterized by the localizedaccumulation of amyloid fibrils in the pancreas.

Once these amyloids have formed, there is no known, widely acceptedtherapy or treatment which significantly dissolves amyloid deposits insitu, prevents further amyloid deposition or prevents the initiation ofamyloid deposition.

Each amyloidogenic protein has the ability to undergo a conformationalchange and to organize into β-sheets and form insoluble fibrils whichmay be deposited extracellularly or intracellularly. Each amyloidogenicprotein, although different in amino acid sequence, has the sameproperty of forming fibrils and binding to other elements such asproteoglycan, amyloid P and complement component. Moreover, eachamyloidogenic protein has amino acid sequences which, althoughdifferent, show similarities such as regions with the ability to bind tothe glycosaminoglycan (GAG) portion of proteoglycan (referred to as theGAG binding site) as well as other regions which promote β-sheetformation. Proteoglycans are macromolecules of various sizes andstructures that are distributed almost everywhere in the body. They canbe found in the intracellular compartment, on the surface of cells, andas part of the extracellular matrix. The basic structure of allproteoglycans is comprised of a core protein and at least one, butfrequently more, polysaccharide chains (GAGs) attached to the coreprotein. Many different GAGs have been discovered including chondroitinsulfate, dermatan sulfate, keratan sulfate, heparin, and hyaluronan.

In specific cases, amyloid fibrils, once deposited, can become toxic tothe surrounding cells. For example, the Aβ fibrils organized as senileplaques have been shown to be associated with dead neuronal cells,dystrophic neurites, astrocytosis, and microgliosis in patients withAlzheimer's disease. When tested in vitro, oligomeric (soluble) as wellas fibrillar Aβ peptide was shown to be capable of triggering anactivation process of microglia (brain macrophages), which would explainthe presence of microgliosis and brain inflammation found in the brainof patients with Alzheimer's disease. Both oligomeric and fibrillar Aβpeptide can also induce neuronal cell death in vitro. See, e.g., M PLambert, et al., Proc. Natl. Acad. Sci. USA 95, 6448-53 (1998).

In another type of amyloidosis seen in patients with type II diabetes,the amyloidogenic protein IAPP, when organized in oligomeric forms or infibrils, has been shown to induce β-islet cell toxicity in vitro. Hence,appearance of IAPP fibrils in the pancreas of type II diabetic patientscontributes to the loss of the β islet cells (Langerhans) and organdysfunction which can lead to insulinemia.

Another type of amyloidosis is related to β₂ microglobulin and is foundin long-term hemodialysis patients. Patients undergoing long termhemodialysis will develop β₂-microglobulin fibrils in the carpal tunneland in the collagen rich tissues in several joints. This causes severepains, joint stiffness and swelling.

Amyloidosis is also characteristic of Alzheimer's disease. Alzheimer'sdisease is a devastating disease of the brain that results inprogressive memory loss leading to dementia, physical disability, anddeath over a relatively long period of time. With the aging populationsin developed countries, the number of Alzheimer's patients is reachingepidemic proportions.

People suffering from Alzheimer's disease develop a progressive dementiain adulthood, accompanied by three main structural changes in the brain:diffuse loss of neurons in multiple parts of the brain; accumulation ofintracellular protein deposits termed neurofibrillary tangles; andaccumulation of extracellular protein deposits termed amyloid or senileplaques, surrounded by misshapen nerve terminals (dystrophic neurites)and activated microglia (microgliosis and astrocytosis). A mainconstituent of these amyloid plaques is the amyloid-β peptide (Aβ), a39-43 amino-acid protein that is produced through cleavage of theβ-amyloid precursor protein (APP). Extensive research has been conductedon the relevance of Aβ deposits in Alzheimer's disease, see, e.g.,Selkoe, Trends in Cell Biology 8, 447-453 (1998). Aβ naturally arisesfrom the metabolic processing of the amyloid precursor protein (“APP”)in the endoplasmic reticulum (“ER”), the Golgi apparatus, or theendosomal-lysosomal pathway, and most is normally secreted as a 40(“Aβ1-40”) or 42 (“Aβ1-42”) amino acid peptide (Selkoe, Annu. Rev. CellBiol. 10, 373-403 (1994)). A role for Aβ as a primary cause forAlzheimer's disease is supported by the presence of extracellular Aβdeposits in senile plaques of Alzheimer's disease, the increasedproduction of Aβ in cells harboring mutant Alzheimer's diseaseassociated genes, e.g., amyloid precursor protein, presenilin I andpresenilin II; and the toxicity of extracellular soluble (oligomeric) orfibrillar Aβ to cells in culture. See, e.g., Gervais, Eur. Biopharm.Review, 40-42 (Autumn 2001); May, DDT 6, 459-62 (2001). Althoughsymptomatic treatments exist for Alzheimer's disease, this diseasecannot be prevented or cured at this time.

Alzheimer's disease is characterized by diffuse and neuritic plaques,cerebral angiopathy, and neurofibrillary tangles. Plaque and bloodvessel amyloid is believed to be formed by the deposition of insolubleAβ amyloid protein, which may be described as diffuse or fibrillary.Both soluble oligomeric Aβ and fibrillar Aβ are also believed to beneurotoxic and inflammatory.

Another type of amyloidosis is cerebral amyloid angiopathy (CAA). CAA isthe specific deposition of amyloid-β fibrils in the walls ofleptomingeal and cortical arteries, arterioles and veins. It is commonlyassociated with Alzheimer's disease, Down's syndrome and normal aging,as well as with a variety of familial conditions related to stroke ordementia (see Frangione et al., Amyloid: J. Protein Folding Disord. 8,Suppl. 1, 36-42 (2001)).

Presently available therapies for treatment of β-amyloid diseases arealmost entirely symptomatic, providing only temporary or partialclinical benefit. Although some pharmaceutical agents have beendescribed that offer partial symptomatic relief, no comprehensivepharmacological therapy is currently available for the prevention ortreatment of, for example, Alzheimer's disease.

SUMMARY OF THE INVENTION

The present invention relates to the use of certain compounds in thetreatment of amyloid-related diseases. In particular, the inventionrelates to a method of treating or preventing an amyloid-related diseasein a subject comprising administering to the subject a therapeuticamount of a compound of the invention. The invention also pertains toeach of the novel compounds of the invention as described herein. Amongthe compounds for use in the invention are those according to thefollowing Formulae, such that, when administered, amyloid fibrilformation, organ specific dysfunction (e.g., neurodegeneration), orcellular toxicity is reduced or inhibited.

In one embodiment, the invention pertains, at least in part to compoundsof Formula I:

wherein:

R¹ is a substituted or unsubstituted cycloalkyl, heterocyclic, aryl,arylcycloalkyl, bicyclic or tricyclic ring, a bicyclic or tricyclicfused ring group, or a substituted or unsubstituted C₂-C₁₀ alkyl group;

R² is selected from a group consisting of hydrogen, alkyl,mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl,triazolyl, imidazolyl, benzothiazolyl, and benzoimidazolyl;

Y is SO₃ ⁻X⁺, OSO₃ ⁻X⁺, or SSO₃ ⁻X⁺;

X⁺ is hydrogen, a cationic group, or an ester-forming group (i.e., as ina prodrug, which are described elsewhere herein); and

each of L¹ and L² is independently a substituted or unsubstituted C₁-C₅alkyl group or absent, or a pharmaceutically acceptable salt thereof,provided that when R¹ is alkyl, L¹ is absent.

In another embodiment, the invention pertains, at least in part tocompounds of Formula II:

wherein:

R¹ is a substituted or unsubstituted cyclic, bicyclic, tricyclic, orbenzoheterocyclic group or a substituted or unsubstituted C₂-C₁₀ alkylgroup;

R² is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl,aryl, arylalkyl, thiazolyl, triazolyl, imidazolyl, benzothiazolyl,benzoimidazolyl, or linked to R¹ to form a heterocycle;

Y is SO₃ ⁻X⁺, OSO₃ ⁻X⁺, or SSO₃ ⁻X⁺;

X⁺ is hydrogen, a cationic group, or an ester forming moiety;

m is 0 or 1;

n is 1, 2, 3, or 4;

L is substituted or unsubstituted C₁-C₃ alkyl group or absent,

or a pharmaceutically acceptable salt thereof, provided that when R¹ isalkyl, L is absent.

In yet another embodiment, the invention pertains, at least in part tocompounds of Formula III:

wherein:

A is nitrogen or oxygen;

R¹¹ is hydrogen, salt-forming cation, ester forming group, —(CH₂)_(x)-Q,or when A is nitrogen, A and R¹¹ taken together may be the residue of anatural or unnatural amino acid or a salt or ester thereof;

Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, orbenzoimidazolyl;

x is 0, 1, 2, 3, or 4;

n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

R³, R^(3a), R⁴, R^(4a), R⁵, R^(5a), R⁶, R^(6a), R⁷ and R^(7a) are eachindependently hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl,cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, cyano,halogen, amino, tetrazolyl, or two R groups on adjacent ring atoms takentogether with the ring atoms form a double bond, provided that one ofR³, R^(3a), R⁴, R^(4a), R⁵, R^(5a), R⁶, R^(6a), R⁷ and R^(7a) is amoiety of Formula IIIa:

wherein:

m is 0, 1, 2, 3, or 4;

R^(A′), R^(B′), R^(C′), R^(D′), and R^(E′) are independently selectedfrom a group of hydrogen, halogen, hydroxyl, alkyl, alkoxyl, halogenatedalkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, cyano,thiazolyl, triazolyl, imidazolyl, tetrazolyl, benzothiazolyl, andbenzoimidazolyl; and pharmaceutically acceptable salts and estersthereof, provided that said compound is not3-(4-phenyl-1,2,3,6-tetrahydro-1-pyridyl)-1-propanesulfonic acid.

In yet another embodiment, the invention pertains at least in part tocompounds of Formula IV:

wherein:

A is nitrogen or oxygen;

R¹¹ is hydrogen, salt-forming cation, ester forming group, —(CH₂)_(x)-Q,or when A is nitrogen, A and R¹¹ taken together may be the residue of anatural or unnatural amino acid or a salt or ester thereof;

Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, orbenzoimidazolyl;

x is 0, 1, 2, 3, or 4;

n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

R⁴, R^(4a), R⁵, R^(5a), R⁶, R^(6a), R⁷, and R^(7a) are eachindependently hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl,cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, cyano,halogen, amino, tetrazolyl, R⁴ and R⁵ taken together, with the ringatoms they are attached to, form a double bond, or R⁶ and R⁷ takentogether, with the ring atoms they are attached to, form a double bond;

m is 0, 1, 2, 3, or 4;

R⁸, R⁹, R¹⁰, R¹¹, and R¹² are independently selected from a group ofhydrogen, halogen, hydroxyl, alkyl, alkoxyl, halogenated alkyl,mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, cyano, thiazolyl,triazolyl, imidazolyl, tetrazolyl, benzothiazolyl, and benzoimidazolyl,and pharmaceutically acceptable salts and esters thereof.

In another embodiment, the invention includes compounds of Formula V:

wherein:

A is nitrogen or oxygen;

R¹¹ is hydrogen, salt-forming cation, ester forming group, —(CH₂)_(x)-Q,or when A is nitrogen, A and R¹¹ taken together may be the residue of anatural or unnatural amino acid or a salt or ester thereof;

Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, orbenzoimidazolyl;

x is 0, 1, 2, 3, or 4;

n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

aa is a natural or unnatural amino acid residue;

m is 0, 1, 2, or 3;

R¹⁴ is hydrogen or protecting group;

R¹⁵ is hydrogen, alkyl or aryl, and pharmaceutically acceptable saltsand prodrugs thereof.

In another embodiment, the invention includes compounds of the FormulaVI:

wherein:

n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

A is oxygen or nitrogen;

R¹¹ is hydrogen, salt-forming cation, ester forming group, —(CH₂)_(x)-Q,or when A is nitrogen, A and R¹¹ taken together may be the residue of anatural or unnatural amino acid or a salt or ester thereof;

Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, orbenzoimidazolyl;

x is 0, 1, 2, 3, or 4;

R¹⁹ is hydrogen, alkyl or aryl;

Y¹ is oxygen, sulfur, or nitrogen;

Y² is carbon, nitrogen, or oxygen;

R²⁰ is hydrogen, alkyl, amino, mercaptoalkyl, alkenyl, alkynyl,cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl,imidazolyl, benzothiazolyl, or benzoimidazolyl;

R²¹ is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl,aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl, imidazolyl,benzothiazolyl, benzoimidazolyl, or absent if Y² is oxygen;

R²² is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl,aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl, imidazolyl,benzothiazolyl, benzoimidazolyl; or R²² is hydrogen, hydroxyl, alkoxy oraryloxy if Y¹ is nitrogen; or R²² is absent if Y¹ is oxygen or sulfur;or R²² and R²¹ may be linked to form a cyclic moiety if Y¹ is nitrogen;

R²³ is hydrogen, alkyl, amino, mercaptoalkyl, alkenyl, alkynyl,cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl,imidazolyl, benzothiazolyl, or benzoimidazolyl, or absent if Y² isnitrogen or oxygen;

or pharmaceutically acceptable salts thereof.

In another embodiment, the invention includes compounds of Formula VII:

wherein:

n is 2, 3, or 4;

A is oxygen or nitrogen;

R¹¹ is hydrogen, salt-forming cation, ester forming group, —(CH₂)_(x)-Q,or when A is nitrogen, A and R¹¹ taken together may be the residue of anatural or unnatural amino acid or a salt or ester thereof;

Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, orbenzoimidazolyl;

x is 0, 1, 2, 3, or 4;

G is a direct bond or oxygen, nitrogen, or sulfur;

z is 0, 1, 2, 3, 4, or 5;

m is 0 or 1;

R²⁴ is selected from a group consisting of hydrogen, alkyl,mercaptoalkyl, alkenyl, alkynyl, aroyl, alkylcarbonyl,aminoalkylcarbonyl, cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl,imidazolyl, benzothiazolyl, and benzoimidazolyl;

each R²⁵ is independently selected from hydrogen, halogen, cyano,hydroxyl, alkoxy, thiol, amino, nitro, alkyl, aryl, carbocyclic, orheterocyclic, and pharmaceutically acceptable salts thereof.

In a further embodiment, the compounds of the invention includecompounds of the formula:

wherein:

R¹ is hydrogen, a substituted or unsubstituted cycloalkyl, heterocyclic,aryl, arylcycloalkyl, bicyclic or tricyclic ring, a bicyclic ortricyclic fused ring group, or a substituted or unsubstituted C₂-C₁₀alkyl group;

R² is selected from a group consisting of hydrogen, alkyl,mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl,triazolyl, imidazolyl, benzothiazolyl, and benzoimidazolyl;

Y is SO₃ ⁻X⁺, OSO₃ ⁻X⁺, or SSO₃ ^(−X) ⁺;

X⁺ is hydrogen, a cationic group, or an ester-forming group;

L¹ is a substituted or unsubstituted C₁-C₅ alkyl group or absent,

B is C₁-C₅ alkyl, alkenyl, or alkynyl group, optionally fused with Wwhen M is absent;

M is a covalent bond, amino, C₁-C₆ alkyl, alkenyl, alkynyl, carboxyl,oxy, amide, ester, thioether, thioester or absent;

W is a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl,aryl, arylalkyl, bicyclic or tricyclic ring, a bicyclic or tricyclicfused ring group, heterocyclic, thiazolyl, triazolyl, imidazolyl,benzothiazolyl, or benzoimidazolyl; and

v is 1, 2, 3, 4, 5, or 6; or a pharmaceutically acceptable salt, esteror prodrug thereof, provided that when Y is methyl, R¹ and R² arehydrogen, Y is SO₃ ⁻X⁺, M is a covalent bond, B is not CH₂—CH(M-W)—CH₂.

In another embodiment, the invention pertains, at least in part, tocompounds of Formula IX:

wherein:

R¹ is a substituted or unsubstituted cycloalkyl, heterocyclic, aryl,arylcycloalkyl, bicyclic or tricyclic ring, a bicyclic or tricyclicfused ring group, or a substituted or unsubstituted C₂-C₁₀ alkyl group;

R² is selected from the group consisting of hydrogen, alkyl,mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl,triazolyl, imidazolyl, benzothiazolyl, and benzoimidazolyl;

R³ is hydrogen or a protecting group;

aa is a natural or unnatural amino acid residue;

L³ is a covalent bond, amino, C₁-C₆ alkyl, alkenyl, alkynyl, carboxyl,amide, aminoalkyl, ether, ester, thioether, thioester or absent;

Y is SO₃ ⁻X⁺, OSO₃ ⁻X⁺, or SSO₃ ⁻X⁺;

X⁺ is hydrogen, a cationic group, or ester-forming group; and

-   -   each of L¹ and L² is independently a substituted or        unsubstituted C₁-C₅ alkyl group or absent, or a pharmaceutically        acceptable salt, ester or prodrug thereof.

In another embodiment, the invention pertains, at least in part, tocompounds of the formula (X):

wherein:

R^(a) is hydrogen, substituted or unsubstituted alkyl, aryl, heteroaryl,carboxyl, alkyloxycarbonyl, or aminocarbonyl;

R^(b) and R^(c) are each selected independently from hydrogen,substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, CONH₂,or R^(b), R^(c) and the carbon atom they are attached to can form asubstituted or unsubstituted cyclic structure of 4 to 12-membered ringor a fused ring system; and

X⁺ is hydrogen, a cationic group, or an ester-forming group, or apharmaceutically acceptable salt, ester, or prodrug thereof.

In another embodiment, the invention also pertains, at least in part, tocompounds of the formula (XI):

wherein:

R^(d) is H or alkyl;

R^(e) and R^(f) are each independently hydrogen, C₁-C₆ alkyl, or R^(e)and R^(f), taken together with the carbon they are attached to, form a 3to 12-membered ring;

R^(g) is independently selected for each occurrence from the groupconsisting of: hydrogen, alkyl, alkoxy, halogen, NO₂, and alkyl-SO₂;

q is 1, 2, 3, 4, or 5;

X⁺ is hydrogen, a cationic group, or an ester-forming group;

Ar is aryl or heteroaryl; and

Z is —(CH₂)₀₋₃—, —(CHOH)—, (CH₂)₁₋₃O(CH₂)₁₋₃, or a carbonyl group, or apharmaceutically acceptable salt, ester, or prodrug thereof.

In another embodiment, the invention also pertains to compounds of theformula (XII):

wherein:

R^(h) is hydrogen, benzyl, aryl-alkyl, aryl, or alkyl;

R^(i), R^(j), R^(k), R^(m), R^(n), and R^(o) are each independentlyhydrogen, substituted or unsubstituted aryl, substituted orunsubstituted benzyl, alkyl, alkenyl, carbocyclic, heterocyclic, absentor together may be linked to form a ring structure;

X⁺ is hydrogen, a cationic group, or an ester-forming group; and

t¹ and t² are each single or double bonds, provided that both t¹ and t²are not both double bonds, or a pharmaceutically acceptable salt, ester,or prodrug thereof.

In another embodiment, the invention also pertains to compounds of theformula (XIII):

wherein:

n¹ is 0, 1, 2, or 3;

P is a covalent bond, alkyl, alkyloxy, amino, alkylamino, sulfur, oralkylthio;

X⁺ is hydrogen, a cationic group, or an ester-forming group; and

R^(p) is a natural or unnatural amino acid residue, or apharmaceutically acceptable salt, ester, or prodrug thereof.

In another embodiment, the invention includes compounds of the formula(XIV):

wherein:

n² is 0, 1, 2, or 3, selected such that three carbons are between theSO₃ ⁻X⁺ group and the nitrogen atom in the ring;

X⁺ is hydrogen, a cationic group, or an ester-forming group;

R^(s) is hydrogen or when n² is 3, R^(s) is (CH₂)₃—SO₃ ⁻X⁺;

R^(q) and R^(r) are each selected independently from hydrogen or alkyl,or a pharmaceutically acceptable salt, ester, or prodrug thereof.

In yet another embodiment, the invention also pertains, at least inpart, to compounds of the formula (XV):

wherein:

R^(t) is hydrogen, alkyl, or aryl;

R^(u) and R^(v) are each independently for each occurrence selected fromhydrogen, aryl, benzyl, alkyl, alkenyl, carbocyclic, heterocyclic, ortwo R^(u) or R^(v) groups on adjacent carbon atoms may form a doublebond, or together with the carbon atoms they are attached to form acarbocyclic or heterocyclic ring;

n³ is 4, 5, 6, or 7; and

X⁺ is hydrogen, a cationic group, or an ester-forming group; or apharmaceutically acceptable salt, ester, or prodrug thereof.

In one embodiment, the compounds disclosed herein prevent or inhibitamyloid protein assembly into insoluble fibrils which, in vivo, aredeposited in various organs, or it favors clearance of pre-formeddeposits or slows deposition in patients already having deposits. Inanother embodiment, the compound may also prevent the amyloid protein,in its soluble, oligomeric form or in its fibrillar form, from bindingor adhering to a cell surface and causing cell damage or toxicity. Inyet another embodiment, the compound may block amyloid-induced cellulartoxicity or macrophage activation. In another embodiment, the compoundmay block amyloid-induced neurotoxicity or microglial activation. Inanother embodiment, the compound protects cells from amyloid inducedcytotoxicity of B-islet cells. In another embodiment, the compound mayenhance clearance from a specific organ, e.g., the brain or it decreasesconcentration of the amyloid protein in such a way that amyloid fibrilformation is prevented in the targeted organ.

The compounds of the invention may be administered therapeutically orprophylactically to treat diseases associated with amyloid fibrilformation, aggregation or deposition. The compounds of the invention mayact to ameliorate the course of an amyloid related disease using any ofthe following mechanisms (this list is meant to be illustrative and notlimiting): slowing the rate of amyloid fibril formation or deposition;lessening the degree of amyloid deposition; inhibiting, reducing, orpreventing amyloid fibril formation; inhibiting neurodegeneration orcellular toxicity induced by amyloid; inhibiting amyloid inducedinflammation; enhancing the clearance of amyloid; or favoring thedegradation of amyloid protein prior to its organization in fibrils.

The compounds of the invention may be administered therapeutically orprophylactically to treat diseases associated with amyloid-β fibrilformation, aggregation or deposition. The compounds of the invention mayact to ameliorate the course of an amyloid-β related disease using anyof the following mechanisms (this list is meant to be illustrative andnot limiting): slowing the rate of amyloid-β fibril formation ordeposition; lessening the degree of amyloid-β deposition; inhibiting,reducing, or preventing amyloid-β fibril formation; inhibitingneurodegeneration or cellular toxicity induced by amyloid-β; inhibitingamyloid-β induced inflammation; enhancing the clearance of amyloid-βfrom the brain; or favoring the degradation of amyloid-β protein priorto its organization in fibrils.

Therapeutic compounds of the invention may be effective in controllingamyloid-β deposition either following their entry into the brain(following penetration of the blood brain barrier) or from theperiphery. When acting from the periphery, a compound may alter theequilibrium of Aβ between the brain and the plasma so as to favor theexit of Aβ from the brain. It may also increase the catabolism ofneuronal Aβ and change the rate of exit from the brain. An increase inthe exit of Aβ from the brain would result in a decrease in Aβ brain andcerebral spinal fluid (CSF) concentration and therefore favor a decreasein Aβ deposition. Alternatively, compounds that penetrate the braincould control deposition by acting directly on brain Aβ e.g., bymaintaining it in a non-fibrillar form, favoring its clearance from thebrain, or by slowing down APP processing. These compounds could alsoprevent Aβ in the brain from interacting with the cell surface andtherefore prevent neurotoxicity, neurodegeneration or inflammation. Theymay also decrease Aβ production by activated microglia. The compoundsmay also increase degradation by macrophages or neuronal cells.

In one embodiment, the method is used to treat Alzheimer's disease(e.g., sporadic, familial, or early AD). The method can also be usedprophylactically or therapeutically to treat other clinical occurrencesof amyloid-β deposition, such as in Down's syndrome individuals and inpatients with cerebral amyloid angiopathy (“CAA”) or hereditary cerebralhemorrhage.

In another embodiment, the method is used to treat mild cognitiveimpairment. Mild Cognitive Impairment (“MCI”) is a conditioncharacterized by a state of mild but measurable impairment in thinkingskills, which is not necessarily associated with the presence ofdementia. MCI frequently, but not necessarily, precedes Alzheimer'sdisease.

Additionally, abnormal accumulation of APP and of amyloid-β protein inmuscle fibers has been implicated in the pathology of sporadic inclusionbody myositis (IBM) (Askanas, et al., Proc. Natl. Acad. Sci. USA 93,1314-1319 (1996); Askanas, et al., Current Opinion in Rheumatology 7,486-496 (1995)). Accordingly, the compounds of the invention can be usedprophylactically or therapeutically in the treatment of disorders inwhich amyloid-beta protein is abnormally deposited at non-neurologicallocations, such as treatment of IBM by delivery of the compounds tomuscle fibers.

Additionally, it has been shown that Aβ is associated with abnormalextracellular deposits, known as drusen, that accumulate along the basalsurface of the retinal pigmented epithelium in individuals withage-related macular degeneration (AMD). AMD is a cause of irreversiblevision loss in older individuals. It is believed that Aβ depositioncould be an important component of the local inflammatory events thatcontribute to atrophy of the retinal pigmented epithelium, drusenbiogenesis, and the pathogenesis of AMD (Johnson, et al., Proc. Natl.Acad. Sci. USA 99(18), 11830-5 (2002)).

The present invention therefore relates to the use of compounds ofFormulae I-XV or otherwise described herein in the prevention ortreatment of amyloid-related diseases, including, inter alia,Alzheimer's disease, cerebral amyloid angiopathy, mild cognitiveimpairment, inclusion body myositis, Down's syndrome, maculardegeneration, as well as other types of amyloidosis such as IAPP-relatedamyloidosis (e.g., diabetes), primary (AL) amyloidosis, secondary (AA)amyloidosis and β₂ microglobulin-related (dialysis-related) amyloidosis.

In Type II diabetes-related amyloidosis (IAPP), the amyloidogenicprotein IAPP induces β-islet cell toxicity when organized in oligomericforms or in fibrils. Hence, appearance of IAPP fibrils in the pancreasof type II diabetic patients contributes to the loss of the β isletcells (Langerhans) and organ dysfunction which leads to insulinemia.

Primary amyloidosis (AL amyloid) is usually found associated with plasmacell dyscrasia and multiple myeloma. It can also be found as anidiopathic disease.

Secondary (AA) amyloidosis is usually seen associated with chronicinfection (such as tuberculosis) or chronic inflammation (such asrheumatoid arthritis). A familial form of secondary amyloidosis is alsoseen in Familial Mediterranean Fever (FMF).

β₂ microglobulin-related (dialysis-related) amyloidosis is found inlong-term hemodialysis patients. Patients undergoing long termhemodialysis will develop β₂-microglobulin fibrils in the carpal tunneland in the collagen rich tissues in several joints. This causes severepains, joint stiffness and swelling. These deposits are due to theinability to maintain low levels of β₂M in plasma of dialyzed patients.Increased plasma concentrations of β₂M protein will induce structuralchanges and may lead to the deposition of modified β₂M as insolublefibrils in the joints.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the use of compounds of Formulae I-XVor compounds otherwise described herein in the treatment ofamyloid-related diseases. For convenience, some definitions of termsreferred to herein are set forth below.

Amyloid-Related Diseases AA (Reactive) Amyloidosis

Generally, AA amyloidosis is a manifestation of a number of diseasesthat provoke a sustained acute phase response. Such diseases includechronic inflammatory disorders, chronic local or systemic microbialinfections, and malignant neoplasms. The most common form of reactive orsecondary (AA) amyloidosis is seen as the result of long-standinginflammatory conditions. For example, patients with Rheumatoid Arthritisor Familial Mediterranean Fever (which is a genetic disease) can developAA amyloidosis. The terms “AA amyloidosis” and “secondary (AA)amyloidosis” are used interchangeably.

AA fibrils are generally composed of 8,000 Dalton fragments (AA peptideor protein) formed by proteolytic cleavage of serum amyloid A protein(ApoSAA), a circulating apolipoprotein which is mainly synthesized inhepatocytes in response to such cytokines as IL-1, IL-6 and TNF. Oncesecreted, ApoSAA is complexed with HDL. Deposition of AA fibrils can bewidespread in the body, with a preference for parenchymal organs. Thekidneys are usually a deposition site, and the liver and the spleen mayalso be affected. Deposition is also seen in the heart, gastrointestinaltract, and the skin.

Underlying diseases which can lead to the development of AA amyloidosisinclude, but are not limited to inflammatory diseases, such asrheumatoid arthritis, juvenile chronic arthritis, ankylosingspondylitis, psoriasis, psoriatic arthropathy, Reiter's syndrome, AdultStill's disease, Behcet's syndrome, and Crohn's disease. AA deposits arealso produced as a result of chronic microbial infections, such asleprosy, tuberculosis, bronchiectasis, decubitus ulcers, chronicpyelonephritis, osteomyelitis, and Whipple's disease. Certain malignantneoplasms can also result in AA fibril amyloid deposits. These includesuch conditions as Hodgkin's lymphoma, renal carcinoma, carcinomas ofgut, lung and urogenital tract, basal cell carcinoma, and hairy cellleukemia. Other underlying conditions that may be associated with AAamyloidosis are Castleman's disease and Schnitzler's syndrome.

AL Amyloidoses (Primary Amyloidosis)

AL amyloid deposition is generally associated with almost any dyscrasiaof the B lymphocyte lineage, ranging from malignancy of plasma cells(multiple myeloma) to benign monoclonal gammopathy. At times, thepresence of amyloid deposits may be a primary indicator of theunderlying dyscrasia. AL amyloidosis is also described in detail inCurrent Drug Targets, 2004, 5 159-171.

Fibrils of AL amyloid deposits are composed of monoclonal immunoglobulinlight chains or fragments thereof. More specifically, the fragments arederived from the N-terminal region of the light chain (kappa or lambda)and contain all or part of the variable (V_(L)) domain thereof. Depositsgenerally occur in the mesenchymal tissues, causing peripheral andautonomic neuropathy, carpal tunnel syndrome, macroglossia, restrictivecardiomyopathy, arthropathy of large joints, immune dyscrasias,myelomas, as well as occult dyscrasias. However, it should be noted thatalmost any tissue, particularly visceral organs such as the kidney,liver, spleen and heart, may be involved.

Hereditary Systemic Amyloidoses

There are many forms of hereditary systemic amyloidoses. Although theyare relatively rare conditions, adult onset of symptoms and theirinheritance patterns (usually autosomal dominant) lead to persistence ofsuch disorders in the general population. Generally, the syndromes areattributable to point mutations in the precursor protein leading toproduction of variant amyloidogenic peptides or proteins. Table 1summarizes the fibril composition of exemplary forms of these disorders.

TABLE 1 Fibril Composition of Exemplary Amyloid-Related Diseases GeneticFibril Peptide/Protein Variant Clinical Syndrome ATTR protein fromTransthyretin Met30, many Familial amyloid polyneuropathy (FAP), andfragments others (Mainly peripheral nerves) ATTR protein fromTransthyretin Thr45, Ala60, Cardiac involvement predominant without andfragments Ser84, Met111, neuropathy, familial amyloid Ile122polyneuropathy, senile systemic amyloidosis, Tenosynovium N-terminalfragment of Arg26 Familial amyloid polyneuropathy (FAP), ApolipoproteinA1 (apoAI) (mainly peripheral nerves) N-terminal fragment of Arg26,Ostertag-type, non-neuropathic Apoliproprotein A1 (AapoAI) Arg50, Arg(predominantly visceral involvement) 60, others AapoAII fromApolipoprotein AII Familial amyloidosis Lysozyme (Alys) Thr56, His67Ostertag-type, non-neuropathic (predominantly visceral involvement)Fibrogen alpha chain fragment Leu554, Cranial neuropathy with latticcorneal Val 526 dystrophy Gelsolin fragment (Agel) Asn187, Cranialneuropathy with lattice corneal Tyr187 dystrophy Cystatin C fragment(ACys) Glu68 Hereditary cerebral hemorrhage (cerebral amyloidangiopathy) - Icelandic type β-amyloid protein (Aβ) derived Gln693Hereditary cerebral hemorrhage (cerebral from Amyloid Precursor Protein(APP) amyloid angiopathy) - Dutch type β-amyloid protein (Aβ) derivedIle717, Phe717, Familial Alzheimer's Disease from Amyloid PrecursorProtein (APP) Gly717 β-amyloid protein (Aβ) derived Gln 618 Alzheimer'sdisease, Down's syndrome, from Amyloid Precursor Protein (APP),hereditary cerebral hemorrhage with e.g., bPP 695 amyloidosis, Dutchtype β-amyloid protein (Aβ) derived Asn670, Familial Dementia - probablyAlzheimer's from Amyloid Precursor Protein (APP) Leu671 Disease PrionProtein (PrP, APrP^(SC)) Leu102, Familial Creutzfeldt-Jakob disease;derived from Prp precursor Val167, Gerstmann-Sträussler-Scheinkersyndrome protein (51-91 insert) Asn178, (hereditary spongiformencephalopathies, Lys200 prion diseases) AA derived from Serum amyloidFamilial Mediterranean fever, predominant A protein (ApoSAA) renalinvolvement (autosomal recessive) AA derived from Serum amyloidMuckle-Well's syndrome, nephropathy, A protein (ApoSAA) deafness,urticaria, limb pain Unknown Cardiomyopathy with persistent atrialstandstill Unknown Cutaneous deposits (bullous, papular, pustulodermal)AH amyloid protein, derived Aγ I Myeloma associated amyloidosis fromimmunoglobulin heavy chain (gamma I) ACal amyloid protein from (Pro)Medullary carcinomas of the thyroid (pro)calcitonin calcitonin AANFamyloid protein from Isolated atrial amyloid atrial natriuretic factorApro from Prolactin Prolactinomas Abri/ADan from ABri peptide Britishand Danish familial Dementia Data derived from Tan S Y, Pepys M B.Amyloidosis. Histopathology, 25(5), 403-414 (November 1994), WHO/IUISNomenclature Subcommittee, Nomenclature of Amyloid and Amyloidosis.Bulletin of the World Health Organisation 1993; 71: 10508; and Merliniet al., Clin Chem Lab Med 2001; 39(11): 1065-75.

The data provided in Table 1 are exemplary and are not intended to limitthe scope of the invention. For example, more than 40 separate pointmutations in the transthyretin gene have been described, all of whichgive rise to clinically similar forms of familial amyloidpolyneuropathy.

In general, any hereditary amyloid disorder can also occur sporadically,and both hereditary and sporadic forms of a disease present with thesame characteristics with regard to amyloid. For example, the mostprevalent form of secondary AA amyloidosis occurs sporadically, e.g. asa result of ongoing inflammation, and is not associated with FamilialMediterranean Fever. Thus general discussion relating to hereditaryamyloid disorders below can also be applied to sporadic amyloidoses.

Transthyretin (TTR) is a 14 kiloDalton protein that is also sometimesreferred to as prealbumin. It is produced by the liver and choroidplexus, and it functions in transporting thyroid hormones and vitamin A.At least 50 variant forms of the protein, each characterized by a singleamino acid change, are responsible for various forms of familial amyloidpolyneuropathy. For example, substitution of proline for leucine atposition 55 results in a particularly progressive form of neuropathy;substitution of methionine for leucine at position 111 resulted in asevere cardiopathy in Danish patients.

Amyloid deposits isolated from heart tissue of patients with systemicamyloidosis have revealed that the deposits are composed of aheterogeneous mixture of TTR and fragments thereof, collectivelyreferred to as ATTR, the full length sequences of which have beencharacterized. ATTR fibril components can be extracted from such plaquesand their structure and sequence determined according to the methodsknown in the art (e.g., Gustaysson, A., et al., Laboratory Invest. 73:703-708, 1995; Kametani, F., et al., Biochem. Biophys. Res. Commun. 125:622-628, 1984; Pras, M., et al., PNAS 80: 539-42, 1983).

Persons having point mutations in the molecule apolipoprotein Al (e.g.,Gly→Arg26; Trp→Arg50; Leu→Arg60) exhibit a form of amyloidosis(“Östertag type”) characterized by deposits of the proteinapolipoprotein AI or fragments thereof (AApoAI). These patients have lowlevels of high density lipoprotein (HDL) and present with a peripheralneuropathy or renal failure.

A mutation in the alpha chain of the enzyme lysozyme (e.g., Ile→Thr56 orAsp→His57) is the basis of another form of Östertag-type non-neuropathichereditary amyloid reported in English families. Here, fibrils of themutant lysozyme protein (Alys) are deposited, and patients generallyexhibit impaired renal function. This protein, unlike most of thefibril-forming proteins described herein, is usually present in whole(unfragmented) form (Benson, M. D., et al. CIBA Fdn. Symp. 199: 104-131,1996).

Immunoglobulin light chains tend to form aggregates in variousmorphologies, including fibrillar (e.g., AL amyloidosis and AHamyloidosis), granular (e.g., light chain deposition disease (LCDD),heavy chain deposition disease (HCDD), and light-heavy chain depositiondisease (LHCDD)), crystalline (e.g., Acquired Farconi's Syndome), andmicrotubular (e.g., Cryoglobulinemia). AL and AH amyloidosis isindicated by the formation of insoluble fibrils of immunoglobulin lightchains and heavy chain, respectively, and/or their fragments. In ALfibrils, lambda (λ) chains such as λ VI chains (λ6 chains), are found ingreater concentrations than kappa (κ) chains. λIII chains are alsoslightly elevated. Merlini et al., CLIN CHEM LAB MED 39(11):1065-75(2001). Heavy chain amyloidosis (AH) is generally characterized byaggregates of gamma chain amyloid proteins of the IgG1 subclass. Eulitzet al., PROC NATL ACAD SCI USA 87:6542-46 (1990).

Comparison of amyloidogenic to non-amyloidogenic light chains hasrevealed that the former can include replacements or substitutions thatappear to destabilize the folding of the protein and promoteaggregation. AL and LCDD have been distinguished from other amyloiddiseases due to their relatively small population monoclonal lightchains, which are manufactured by neoplastic expansion of anantibody-producing B cell. AL aggregates typically are well-orderedfibrils of lambda chains. LCDD aggregates are relatively amorphousaggregations of both kappa and lambda chains, with a majority beingkappa, in some cases κIV. Bellotti et al., JOURNAL OF STRUCTURAL BIOLOGY13:280-89 (2000). Comparison of amyloidogenic and non-amyloidogenicheavy chains in patients having AH amyloidosis has revealed missingand/or altered components. Eulitz et al., PROC NATL ACAD SCI USA87:6542-46 (1990) (pathogenic heavy chain characterized by significantlylower molecular mass than non-amyloidogenic heavy chains); and Solomonet al. AM J HEMAT 45(2) 171-6 (1994) (amyloidogenic heavy chaincharacterized as consisting solely of the VH-D portion of thenon-amyloidogenic heavy chain).

Accordingly, potential methods of detecting and monitoring treatment ofsubjects having or at risk of having AL, LCDD, AH, and the like, includebut are not limited to immunoassaying plasma or urine for the presenceor depressed deposition of amyloidogenic light or heavy chains, e.g.,amyloid λ, amyloid κ, amyloid κIV, amyloid γ, or amyloid γ1.

Brain Amyloidosis

The most frequent type of amyloid in the brain is composed primarily ofAβ peptide fibrils, resulting in dementia associated with sporadic(non-hereditary) Alzheimer's disease. In fact, the incidence of sporadicAlzheimer's disease greatly exceeds forms shown to be hereditary.Nevertheless, fibril peptides forming plaques are very similar in bothtypes. Brain amyloidosis includes those diseases, conditions,pathologies, and other abnormalities of the structure or function of thebrain, including components thereof, in which the causative agent isamyloid. The area of the brain affected in an amyloid-related diseasemay be the stroma including the vasculature or the parenchyma includingfunctional or anatomical regions, or neurons themselves. A subject neednot have received a definitive diagnosis of a specifically recognizedamyloid-related disease. The term “amyloid related disease” includesbrain amyloidosis.

Amyloid-β peptide (“Aβ”) is a 39-43 amino acid peptide derived byproteolysis from a large protein known as Beta Amyloid Precursor Protein(“βAPP”). Mutations in βAPP result in familial forms of Alzheimer'sdisease, Down's syndrome, cerebral amyloid angiopathy, and seniledementia, characterized by cerebral deposition of plaques composed of Aβfibrils and other components, which are described in further detailbelow. Known mutations in APP associated with Alzheimer's disease occurproximate to the cleavage sites of β or γ-secretase, or within Aβ. Forexample, position 717 is proximate to the site of gamma-secretasecleavage of APP in its processing to Aβ, and positions 670/671 areproximate to the site of β-secretase cleavage. Mutations at any of theseresidues may result in Alzheimer's disease, presumably by causing anincrease in the amount of the 42/43 amino acid form of Aβ generated fromAPP. The familial form of Alzheimer's disease represents only 10% of thesubject population. Most occurrences of Alzheimer's disease are sporadiccases where APP and Aβ do not possess any mutation. The structure andsequence of Aβ peptides of various lengths are well known in the art.Such peptides can be made according to methods known in the art, orextracted from the brain according to known methods (e.g., Glenner andWong, Biochem. Biophys. Res. Comm. 129, 885-90 (1984); Glenner and Wong,Biochem. Biophys. Res. Comm. 122, 1131-35 (1984)). In addition, variousforms of the peptides are commercially available. APP is expressed andconstitutively catabolized in most cells. The dominant catabolic pathwayappears to be cleavage of APP within the Aβ sequence by an enzymeprovisionally termed α-secretase, leading to release of a solubleectodomain fragment known as APPsα. This cleavage precludes theformation of Aβ peptide. In contrast to this non-amyloidogenic pathway,APP can also be cleaved by enzymes known as β- and γ-secretase at the N-and C-termini of the Aβ, respectively, followed by release of Aβ intothe extracellular space. To date, BACE has been identified asβ-secretase (Vasser, et al., Science 286:735-741, 1999) and presenilinshave been implicated in γ-secretase activity (De Strooper, et al.,Nature 391, 387-90 (1998)). The 39-43 amino acid Aβ peptide is producedby sequential proteolytic cleavage of the amyloid precursor protein(APP) by the β and γ secretases enzyme. Although Aβ40 is the predominantform produced, 5-7% of total Aβ exists as Aβ42 (Cappai et al., Int. J.Biochem. Cell Biol. 31. 885-89 (1999)).

The length of the Aβ peptide appears to dramatically alter itsbiochemical/biophysical properties. Specifically, the additional twoamino acids at the C-terminus of Aβ42 are very hydrophobic, presumablyincreasing the propensity of Aβ42 to aggregate. For example, Jarrett, etal. demonstrated that Aβ42 aggregates very rapidly in vitro compared toAβ40, suggesting that the longer forms of Aβ may be the importantpathological proteins that are involved in the initial seeding of theneuritic plaques in Alzheimer's disease (Jarrett, et al., Biochemistry32, 4693-97 (1993); Jarrett, et al., Ann. N.Y. Acad. Sci. 695, 144-48(1993)). This hypothesis has been further substantiated by the recentanalysis of the contributions of specific forms of Aβ in cases ofgenetic familial forms of Alzheimer's disease (“FAD”). For example, the“London” mutant form of APP (APPV717I) linked to FAD selectivelyincreases the production of Aβ 42/43 forms versus Aβ 40 (Suzuki, et al.,Science 264, 1336-40 (1994)) while the “Swedish” mutant form of APP(APPK670N/M671L) increases levels of both Aβ40 and Aβ42/43 (Citron, etal., Nature 360, 672-674 (1992); Cai, et al., Science 259, 514-16,(1993)). Also, it has been observed that FAD-linked mutations in thePresenilin-1 (“PS1”) or Presenilin-2 (“PS2”) genes will lead to aselective increase in Aβ42/43 production but not Aβ40 (Borchelt, et al.,Neuron 17, 1005-13 (1996)). This finding was corroborated in transgenicmouse models expressing PS mutants that demonstrate a selective increasein brain Aβ42 (Borchelt, op cit.; Duff, et al., Neurodegeneration 5(4),293-98 (1996)). Thus the leading hypothesis regarding the etiology ofAlzheimer's disease is that an increase in Aβ42 brain concentration dueto an increased production and release of Aβ42 or a decrease inclearance (degradation or brain clearance) is a causative event in thedisease pathology.

Multiple mutation sites in either Aβ or the APP gene have beenidentified and are clinically associated with either dementia orcerebral hemorrhage. Exemplary CAA disorders include, but are notlimited to, hereditary cerebral hemorrhage with amyloidosis of Icelandictype (HCHWA-I); the Dutch variant of HCHWA (HCHWA-D; a mutation in Aβ);the Flemish mutation of Aβ; the Arctic mutation of Aβ; the Italianmutation of Aβ; the Iowa mutation of Aβ; familial British dementia; andfamilial Danish dementia. CAA may also be sporadic.

As used herein, the terms “β amyloid,” “amyloid-β,” and the like referto amyloid β proteins or peptides, amyloid β precursor proteins orpeptides, intermediates, and modifications and fragments thereof, unlessotherwise specifically indicated. In particular, “Aβ” refers to anypeptide produced by proteolytic processing of the APP gene product,especially peptides which are associated with amyloid pathologies,including Aβ1-39, Aβ1-40, Aβ1-41, Aβ1-42, and Aβ1-43. For convenience ofnomenclature, “Aβ1-42” may be referred to herein as “Aβ(1-42)” or simplyas “Aβ42” or “Aβ₄₂” (and likewise for any other amyloid peptidesdiscussed herein). As used herein, the terms “β amyloid,” “amyloid-β,”and “Aβ” are synonymous.

Unless otherwise specified, the term “amyloid” refers to amyloidogenicproteins, peptides, or fragments thereof which can be soluble (e.g.,monomeric or oligomeric) or insoluble (e.g., having fibrillary structureor in amyloid plaque). See, e.g., M P Lambert, et al., Proc. Nat'l Acad.Sci. USA 95, 6448-53 (1998). “Amyloidosis” or “amyloid disease” or“amyloid-related disease” refers to a pathological conditioncharacterized by the presence of amyloid fibers. “Amyloid” is a genericterm referring to a group of diverse but specific protein deposits(intracellular or extracellular) which are seen in a number of differentdiseases. Though diverse in their occurrence, all amyloid deposits havecommon morphologic properties, stain with specific dyes (e.g., Congored), and have a characteristic red-green birefringent appearance inpolarized light after staining. They also share common ultrastructuralfeatures and common X-ray diffraction and infrared spectra.

Gelsolin is a calcium binding protein that binds to fragments and actinfilaments. Mutations at position 187 (e.g., Asp→Asn; Asp→Tyr) of theprotein result in a form of hereditary systemic amyloidosis, usuallyfound in patients from Finland, as well as persons of Dutch or Japaneseorigin. In afflicted individuals, fibrils formed from gelsolin fragments(Agel), usually consist of amino acids 173-243 (68 kDa carboxyterminalfragment) and are deposited in blood vessels and basement membranes,resulting in corneal dystrophy and cranial neuropathy which progressesto peripheral neuropathy, dystrophic skin changes and deposition inother organs. (Kangas, H., et al. Human Mol. Genet. 5(9): 1237-1243,1996).

Other mutated proteins, such as mutant alpha chain of fibrinogen (AfibA)and mutant cystatin C (Acys) also form fibrils and producecharacteristic hereditary disorders. AfibA fibrils form depositscharacteristic of a nonneuropathic hereditary amyloid with renaldisease; Acys deposits are characteristic of a hereditary cerebralamyloid angiopathy reported in Iceland (Isselbacher, Harrison'sPrinciples of Internal Medicine, McGraw-Hill, San Francisco, 1995;Benson, et al.). In at least some cases, patients with cerebral amyloidangiopathy (CAA) have been shown to have amyloid fibrils containing anon-mutant form of cystatin C in conjunction with amyloid beta protein(Nagai, A., et al. Molec. Chem. Neuropathol. 33: 63-78, 1998).

Certain forms of prion disease are now considered to be heritable,accounting for up to 15% of cases, which were previously thought to bepredominantly infectious in nature. (Baldwin, et al., in ResearchAdvances in Alzheimer's Disease and Related Disorders, John Wiley andSons, New York, 1995). In hereditary and sporadic prion disorders,patients develop plaques composed of abnormal isoforms of the normalprion protein (PrP^(Sc)).

A predominant mutant isoform, PrP^(Sc), also referred to as AScr,differs from the normal cellular protein in its resistance to proteasedegradation, insolubility after detergent extraction, deposition insecondary lysosomes, post-translational synthesis, and high β-pleatedsheet content. Genetic linkage has been established for at least fivemutations resulting in Creutzfeldt-Jacob disease (CJD), Gerstmann-Sträussler-Scheinker syndrome (GSS), and fatal familial insomnia (FFI).(Baldwin, supra) Methods for extracting fibril peptides from scrapiefibrils, determining sequences and making such peptides are known in theart (e.g., Beekes, M., et al. J. Gen. Virol. 76: 2567-76, 1995).

For example, one form of GSS has been linked to a PrP mutation at codon102, while telencephalic GSS segregates with a mutation at codon 117.Mutations at codons 198 and 217 result in a form of GSS in whichneuritic plaques characteristic of Alzheimer's disease contain PrPinstead of Aβ peptide. Certain forms of familial CJD have beenassociated with mutations at codons 200 and 210; mutations at codons 129and 178 have been found in both familial CJD and FFI. (Baldwin, supra).

Cerebral Amyloidosis

Local deposition of amyloid is common in the brain, particularly inelderly individuals. The most frequent type of amyloid in the brain iscomposed primarily of Aβ peptide fibrils, resulting in dementia orsporadic (non-hereditary) Alzheimer's disease. The most commonoccurrences of cerebral amyloidosis are sporadic and not familial. Forexample, the incidence of sporadic Alzheimer's disease and sporadic CAAgreatly exceeds the incidence of familial AD and CAA. Moreover, sporadicand familial forms of the disease cannot be distinguished from eachother (they differ only in the presence or absence of an inheritedgenetic mutation); for example, the clinical symptoms and the amyloidplaques formed in both sporadic and familial AD are very similar, if notidentical.

Cerebral amyloid angiopathy (CAA) refers to the specific deposition ofamyloid fibrils in the walls of leptomingeal and cortical arteries,arterioles and veins. It is commonly associated with Alzheimer'sdisease, Down's syndrome and normal aging, as well as with a variety offamilial conditions related to stroke or dementia (see Frangione et al.,Amyloid: J. Protein Folding Disord. 8, Suppl. 1, 36-42 (2001)). CAA canoccur sporadically or be hereditary.

Senile Systemic Amyloidosis

Amyloid deposition, either systemic or focal, increases with age. Forexample, fibrils of wild type transthyretin (TTR) are commonly found inthe heart tissue of elderly individuals. These may be asymptomatic,clinically silent, or may result in heart failure. Asymptomaticfibrillar focal deposits may also occur in the brain (Aβ), corporaamylacea of the prostate (β₂ microglobulin), joints and seminalvesicles.

Dialysis-Related Amyloidosis (DRA)

Plaques composed of β₂ microglobulin (β₂M) fibrils commonly develop inpatients receiving long term hemodialysis or peritoneal dialysis. β₂microglobulin is a 11.8 kiloDalton polypeptide and is the light chain ofClass I MHC antigens, which are present on all nucleated cells. Undernormal circumstances, β₂M is usually distributed in the extracellularspace unless there is an impaired renal function, in which case β₂M istransported into tissues where it polymerizes to form amyloid fibrils.Failure of clearance such as in the case of impaired renal function,leads to deposition in the carpal tunnel and other sites (primarily incollagen-rich tissues of the joints). Unlike other fibril proteins, β₂Mmolecules are not produced by cleavage of a longer precursor protein andare generally present in unfragmented form in the fibrils. (Benson,supra). Retention and accumulation of this amyloid precursor has beenshown to be the main pathogenic process underlying DRA. DRA ischaracterized by peripheral joint osteoarthropathy (e.g., jointstiffness, pain, swelling, etc.). Isoforms of β₂M, glycated β₂M, orpolymers of β₂M in tissue are the most amyloidogenic form (as opposed tonative β₂M). Unlike other types of amyloidosis, β₂M is confined largelyto osteoarticular sites. Visceral depositions are rare. Occasionally,these deposits may involve blood vessels and other important anatomicsites.

Despite improved dialysis methods for removal of β₂M, the majority ofpatients have plasmatic β₂M concentrations that remain dramaticallyhigher than normal. These elevated β₂M concentrations generally lead toDiabetes-Related Amyloidosis (DRA) and cormorbidities that contribute tomortality.

Islet Amyloid Polypeptide and Diabetes

Islet hyalinosis (amyloid deposition) was first described over a centuryago as the presence of fibrous protein aggregates in the pancreas ofpatients with severe hyperglycemia (Opie, E L., J Exp. Med. 5: 397-428,1901). Today, islet amyloid, composed predominantly of islet amyloidpolypeptide (IAPP), or amylin, is a characteristic histopathologicalmarker in over 90% of all cases of Type II diabetes (also known asNon-Insulin Dependent Diabetes, or NIDDM). These fibrillar accumulationsresult from the aggregation of the islet amyloid polypeptide (IAPP) oramylin, which is a 37 amino acid peptide, derived from a largerprecursor peptide, called pro-IAPP.

IAPP is co-secreted with insulin in response to β-cell secretagogues.This pathological feature is not associated with insulin-dependent (TypeI) diabetes and is a unifying characteristic for the heterogeneousclinical phenotypes diagnosed as NIDDM (Type II diabetes).

Longitudinal studies in cats and immunocytochemical investigations inmonkeys have shown that a progressive increase in islet amyloid isassociated with a dramatic decrease in the population ofinsulin-secreting β-cells and increased severity of the disease. Morerecently, transgenic studies have strengthened the relationship betweenIAPP plaque formation and β-cell apoptosis and dysfunction, indicatingthat amyloid deposition is a principal factor in increasing severity ofType II diabetes.

IAPP has also been shown to induce β-islet cell toxicity in vitro,indicating that appearance of IAPP fibrils in the pancreas of Type II orType I diabetic patients (post-islet transplantation) could contributeto the loss of the n-cell islets (Langerhans) and organ dysfunction. Inpatients with Type II diabetes, the accumulation of pancreatic IAPPleads to formation of oligomeric IAPP, leading to a buildup ofIAPP-amyloid as insoluble fibrous deposits which eventually destroys theinsulin-producing β cells of the islet, resulting in β cell depletionand failure (Westermark, P., Grimelius, L., Acta Path. Microbiol.Scand., sect. A. 81: 291-300, 1973; de Koning, E J P., et al.,Diabetologia 36: 378-384, 1993; and Lorenzo, A., et al., Nature 368:756-760, 1994). Accumulation of IAPP as fibrous deposits can also havean impact on the ratio of pro-IAPP to IAPP normally found in plasma byincreasing this ratio due to the trapping of IAPP in deposits. Reductionof β cell mass can be manifested by hyperglycemia and insulinemia. Thisβ-cell mass loss can lead to a need for insulin therapy.

Diseases caused by the death or malfunctioning of a particular type ortypes of cells can be treated by transplanting into the patient healthycells of the relevant type of cell. This approach has been used for TypeI diabetes patients. Often pancreatic islet cells from a donor arecultured in vitro prior to transplantation, to allow them to recoverafter the isolation procedure or to reduce their immunogenicity.However, in many instances islet cell transplantation is unsuccessful,due to death of the transplanted cells. One reason for this poor successrate is IAPP, which organizes into toxic oligomers. Toxic effects mayresult from intracellular and extracellular accumulation of fibriloligomers. The IAPP oligomers can form fibrils and become toxic to thecells in vitro. In addition, IAPP fibrils are likely to continue to growafter the cells are transplanted and cause death or dysfunction of thecells. This may occur even when the cells are from a healthy donor andthe patient receiving the transplant does not have a disease that ischaracterized by the presence of fibrils. For example, compounds of thepresent invention may also be used in preparing tissues or cells fortransplantation according to the methods described in InternationalPatent Application (PCT) number WO 01/003680.

The compounds of the invention may also stabilize the ratio of theconcentrations of Pro-IAPP/IAPP, pro-Insulin/Insulin and C-peptidelevels. In addition, as biological markers of efficacy, the results ofthe different tests, such as the arginine-insulin secretion test, theglucose tolerance test, insulin tolerance and sensitivity tests, couldall be used as markers of reduced β-cell mass and/or accumulation ofamyloid deposits. Such class of drugs could be used together with otherdrugs targeting insulin resistance, hepatic glucose production, andinsulin secretion. Such compounds might prevent insulin therapy bypreserving β-cell function and be applicable to preserving islettransplants.

Hormone-Derived Amyloidoses

Endocrine organs may harbor amyloid deposits, particularly in agedindividuals. Hormone-secreting tumors may also contain hormone-derivedamyloid plaques, the fibrils of which are made up of polypeptidehormones such as calcitonin (medullary carcinoma of the thyroid), andatrial natriuretic peptide (isolated atrial amyloidosis). Sequences andstructures of these proteins are well known in the art.

Miscellaneous Amyloidoses

There are a variety of other forms of amyloid disease that are normallymanifest as localized deposits of amyloid. In general, these diseasesare probably the result of the localized production or lack ofcatabolism of specific fibril precursors or a predisposition of aparticular tissue (such as the joint) for fibril deposition. Examples ofsuch idiopathic deposition include nodular AL amyloid, cutaneousamyloid, endocrine amyloid, and tumor-related amyloid. Other amyloidrelated diseases include those described in Table 1, such as familialamyloid polyneuropathy (FAP), senile systemic amyloidosis, Tenosynovium,familial amyloidosis, Ostertag-type, non-neuropathic amyloidosis,cranial neuropathy, hereditary cerebral hemorrhage, familial dementia,chronic dialysis, familial Creutzfeldt-Jakob disease; Gerstmann-Sträussler-Scheinker syndrome, hereditary spongiform encephalopathies, priondiseases, familial Mediterranean fever, Muckle-Well's syndrome,nephropathy, deafness, urticaria, limb pain, cardiomyopathy, cutaneousdeposits, multiple myeloma, benign monoclonal gammopathy,maccoglobulinaemia, myeloma associated amyloidosis, medullary carcinomasof the thyroid, isolated atrial amyloid, and diabetes.

The compounds of the invention may be administered therapeutically orprophylactically to treat diseases associated with amyloid fibrilformation, aggregation or deposition, regardless of the clinicalsetting. The compounds of the invention may act to ameliorate the courseof an amyloid related disease using any of the following mechanisms,such as, for example but not limited to: slowing the rate of amyloidfibril formation or deposition; lessening the degree of amyloiddeposition; inhibiting, reducing, or preventing amyloid fibrilformation; inhibiting amyloid induced inflammation; enhancing theclearance of amyloid from, for example, the brain; or protecting cellsfrom amyloid induced (oligomers or fibrillar) toxicity.

In an embodiment, the compounds of the invention may be administeredtherapeutically or prophylactically to treat diseases associated withamyloid-β fibril formation, aggregation or deposition. The compounds ofthe invention may act to ameliorate the course of an amyloid-β relateddisease using any of the following mechanisms (this list is meant to beillustrative and not limiting): slowing the rate of amyloid-β fibrilformation or deposition; lessening the degree of amyloid-β deposition;inhibiting, reducing, or preventing amyloid-β fibril formation;inhibiting neurodegeneration or cellular toxicity induced by amyloid-β;inhibiting amyloid-β induced inflammation; enhancing the clearance ofamyloid-β from the brain; or favoring greater catabolism of Aβ.

Compounds of the invention may be effective in controlling amyloid-βdeposition either following their entry into the brain (followingpenetration of the blood brain barrier) or from the periphery. Whenacting from the periphery, a compound may alter the equilibrium of Aβbetween the brain and the plasma so as to favor the exit of Aβ from thebrain. An increase in the exit of Aβ from the brain would result in adecrease in Aβ brain concentration and therefore favor a decrease in Aβdeposition. In addition, compounds that penetrate the brain may controldeposition by acting directly on brain Aβ, e.g., by maintaining it in anon-fibrillar form or favoring its clearance from the brain. Thecompounds may slow down APP processing; may increase degradation of Aβfibrils by macrophages or by neuronal cells; or may decrease Aβproduction by activated microglia. These compounds could also prevent Aβin the brain from interacting with the cell surface and thereforeprevent neurotoxicity, neurodegeneration, or inflammation.

In a preferred embodiment, the method is used to treat Alzheimer'sdisease (e.g., sporadic or familial AD). The method can also be usedprophylactically or therapeutically to treat other clinical occurrencesof amyloid-β deposition, such as in Down's syndrome individuals and inpatients with cerebral amyloid angiopathy (“CAA”), hereditary cerebralhemorrhage, or early Alzheimer's disease.

In another embodiment, the method is used to treat mild cognitiveimpairment. Mild Cognitive Impairment (“MCI”) is a conditioncharacterized by a state of mild but measurable impairment in thinkingskills, which is not necessarily associated with the presence ofdementia. MCI frequently, but not necessarily, precedes Alzheimer'sdisease.

Additionally, abnormal accumulation of APP and of amyloid-β protein inmuscle fibers has been implicated in the pathology of sporadic inclusionbody myositis (IBM) (Askanas, V., et al. (1996) Proc. Natl. Acad. Sci.USA 93: 1314-1319; Askanas, V. et al. (1995) Current Opinion inRheumatology 7: 486-496). Accordingly, the compounds of the inventioncan be used prophylactically or therapeutically in the treatment ofdisorders in which amyloid-beta protein is abnormally deposited atnon-neurological locations, such as treatment of IBM by delivery of thecompounds to muscle fibers.

Additionally, it has been shown that Aβ is associated with abnormalextracellular deposits, known as drusen, that accumulate along the basalsurface of the retinal pigmented epithelium in individuals withage-related macular degeneration (ARMD). ARMD is a cause of irreversiblevision loss in older individuals. It is believed that Aβ depositioncould be an important component of the local inflammatory events thatcontribute to atrophy of the retinal pigmented epithelium, drusenbiogenesis, and the pathogenesis of ARMD (Johnson, et al., Proc. Natl.Acad. Sci. USA 99(18), 11830-5 (2002)).

In another embodiment, the invention also relates to a method oftreating or preventing an amyloid-related disease in a subject(preferably a human) comprising administering to the subject atherapeutic amount of a compound according to the following Formulae orotherwise described herein, such that amyloid fibril formation ordeposition, neurodegeneration, or cellular toxicity is reduced orinhibited. In another embodiment, the invention relates to a method oftreating or preventing an amyloid-related disease in a subject(preferably a human) comprising administering to the subject atherapeutic amount of a compound according to the following Formulae orotherwise described herein, such that cognitive function is improved orstabilized or further deterioration in cognitive function is prevented,slowed, or stopped in patients with brain amyloidosis, e.g., Alzheimer'sdisease, Down's syndrome or cerebral amyloid angiopathy. These compoundscan also improve quality of daily living in these subjects.

The therapeutic compounds of the invention may treat amyloidosis relatedto type II diabetes by, for example, stabilizing glycemia, preventing orreducing the loss of β cell mass, reducing or preventing hyperglycemiadue to loss of a cell mass, and modulating (e.g., increasing orstabilizing) insulin production. The compounds of the invention may alsostabilize the ratio of the concentrations of pro-IAPP/IAPP.

The therapeutic compounds of the invention may treat AA (secondary)amyloidosis and/or AL (primary) amyloidosis, by stabilizing renalfunction, decreasing proteinuria, increasing creatinine clearance (e.g.,by at least 50% or greater or by at least 100% or greater), by leadingto remission of chronic diarrhea or weight gain (e.g., 10% or greater),or by reducing serum creatinine. Visceral amyloid content as determined,e.g., by SAP scintigraphy may also be reduced.

Compounds of the Invention

The present invention relates, at least in part, to the use of certainchemical compounds (and pharmaceutical formulations thereof) in theprevention or treatment of amyloid-related diseases, including, interalia, Alzheimer's disease, cerebral amyloid angiopathy, inclusion bodymyositis, Down's syndrome, diabetes related amyloidosis,hemodialysis-related amyloidosis (β₂M), primary amyloidosis (e.g., X orK chain-related), familial amyloid polyneuropathy (FAP), senile systemicamyloidosis, familial amyloidosis, Ostertag-type non-neuropathicamyloidosis, cranial neuropathy, hereditary cerebral hemorrhage,familial dementia, chronic dialysis, familial Creutzfeldt-Jakob disease,Gerstmann-Strä ussler-Scheinker syndrome, hereditary spongiformencephalopathies, prion diseases, familial Mediterranean fever,Muckle-Well's syndrome, nephropathy, deafness, urticaria, limb pain,cardiomyopathy, cutaneous deposits, multiple myeloma, benign monoclonalgammopathy, maccoglobulinaemia, myeloma associated amyloidosis,medullary carcinomas of the thyroid, and isolated atrial amyloid.

The chemical structures herein are drawn according to the conventionalstandards known in the art. Thus, where an atom, such as a carbon atom,as drawn appears to have an unsatisfied valency, then that valency isassumed to be satisfied by a hydrogen atom even though that hydrogenatom is not necessarily explicitly drawn. The structures of some of thecompounds of this invention include stereogenic carbon atoms. It is tobe understood that isomers arising from such asymmetry (e.g., allenantiomers and diastereomers) are included within the scope of thisinvention unless indicated otherwise. That is, unless otherwisestipulated, any chiral carbon center may be of either (R)- or(S)-stereochemistry. Such isomers can be obtained in substantially pureform by classical separation techniques and bystereochemically-controlled synthesis. Furthermore, alkenes can includeeither the E- or Z-geometry, where appropriate. In addition, thecompounds of the present invention may exist in unsolvated as well assolvated forms with acceptable solvents such as water, THF, ethanol, andthe like. In general, the solvated forms are considered equivalent tothe unsolvated forms for the purposes of the present invention.

A “small molecule” refers to a compound that is not itself the productof gene transcription or translation (e.g., protein, RNA, or DNA) andpreferably has a low molecular weight, e.g., less than about 2500 amu.

In general, the term “nucleophile” is art-recognized to mean a chemicalgroup having a reactive pair of electrons that reacts with a compound bydisplacing a leaving group (commonly another nucleophile), such ascommonly occur in aliphatic chemistry as unimolecular (known as“S_(N)1”) or bimolecular (“S_(N)2”) reactions. Examples of nucleophilesinclude uncharged compounds such as amines, mercaptans, and alcohols,and charged groups such as alkoxides, thiolates, carbanions, and avariety of organic and inorganic anions. Illustrative anionicnucleophiles include, inter alia, simple anions such as azide, cyanide,thiocyanate, acetate, formate, or chloroformate, and bisulfite.Organometallic reagents such as organocuprates, organozincs,organolithiums, Grignard reagents, enolates, and acetylides, will underappropriate reaction conditions, be suitable nucleophiles.

Similarly, an “electrophile” means an atom, molecule, or ion able toaccept an electron pair, particularly a pair of electrons from anucleophile, such as typically occurs during an electrophilicsubstitution reaction. In an electrophilic substitution reaction, anelectrophile binds to a substrate with the expulsion of anotherelectrophile, e.g., the substitution of a proton by another electrophilesuch as a nitronium ion on an aromatic substrate (e.g., benzene).Electrophiles include cyclic compounds such as epoxides, aziridines,episulfides, cyclic sulfates, carbonates, lactones, and lactams; andnon-cyclic electrophiles include sulfates, sulfonates (e.g., tosylates),chlorides, bromides, and iodides. Generally, an electrophile may be asaturated carbon atom (e.g., a methylene group) bonded to a leavinggroup; however, an electrophile may also be an unsaturated group, suchas an aldehyde, ketone, ester, or conjugated (α,β-unsaturated) analogthereof, which upon reaction with a nucleophile forms an adduct.

The term “leaving group” generally refers to a group that is readilydisplaced and substituted by a nucleophile (e.g., an amine, a thiol, analcohol, or cyanide). Such leaving groups are well known and includecarboxylates, N-hydroxysuccinimide (“NHS”), N-hydroxybenzotriazole, ahalogen (fluorine, chlorine, bromine, or iodine), alkoxides, andthioalkoxides. A variety of sulfur-based leaving groups are routinelyused in synthetic chemistry, including alkane sulfonyloxy groups (e.g.,C₁-C₄ alkane such as methane sulfonyloxy, ethane sulfonyloxy, propanesulfonyloxy, and butane sulfonyloxy groups) and the halogenated analogs(e.g., halogeno(C₁-C₄ alkane) sulfonyloxy groups, such astrifluoromethane sulfonyloxy (i.e., triflate), 2,2,2-trichloroethanesulfonyloxy, 3,3,3-tribromopropane sulfonyloxy, and4,4,4-trifluorobutane sulfonyloxy groups), as well as arylsulfonyloxygroups (e.g., C₆-C₁₀ aryl optionally substituted with 1 to 3 C₁-C₄ alkylgroups, such as benzene sulfonyloxy, α-naphthylsulfonyloxy,β-naphthylsulfonyloxy, p-toluenesulfonyloxy (i.e., tosylates),4-tert-butylbenzene sulfonyloxy, mesitylene sulfonyloxy, and6-ethyl-α-naphthylsulfonyloxy groups).

“Activated esters” may be represented by the formula —COL, where L is aleaving group, typical examples of which includeN-hydroxysulfosuccinimidyl and N-hydroxysuccinimidyl groups; aryloxygroups substituted with electron-withdrawing groups (e.g., p-nitro,pentafluoro, pentachloro, p-cyano, or p-trifluoromethyl); and carboxylicacids activated by a carbodiimide to form an anhydride or mixedanhydride, e.g., —OCOR^(a) or —OCNR^(a)NHR^(b), where R^(a) and R^(b)are independently C₁-C₆ alkyl, C₅-C₈ alkyl (e.g., cyclohexyl), C₁-C₆perfluoroalkyl, or C₁-C₆ alkoxy groups. An activated ester may be formedin situ or may be an isolable reagent. Sulfosuccinimidyl esters,pentafluorothiophenol esters, and sulfotetrafluorophenol are preferredactivated esters. However, the ester leaving group may be, for example,substituted or unsubstituted C₁-C₆ alkyl (such as methyl, ethyl, propyl,isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, or hexyl), orsubstituted or unsubstituted C₆-C₁₄ aryl or heterocyclic groups, such as2-fluoroethyl, 2-chloroethyl, 2-bromoethyl, 2,2-dibromoethyl,2,2,2-trichloroethyl, 3-fluoropropyl, 4-chlorobutyl, methoxymethyl,1,1-dimethyl-1-methoxymethyl, ethoxymethyl, N-propoxymethyl,isopropoxymethyl, N-butoxymethyl, tert-butoxymethyl, 1-ethoxyethyl,1-methyl-1-methoxyethyl, 1-(isopropoxy)ethyl,3-methoxypropyl-4-methoxybutyl, fluoromethoxymethyl,2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl,3-fluoropropoxymethyl, 4-chlorobutoxyethyl, dibromomethoxyethyl,2-chloroethoxypropyl, fluoromethoxybutyl, 2-methoxyethoxymethyl,ethoxymethoxyethyl, methoxyethoxypropyl, methoxyethoxybutyl, benzyl,phenethyl, 3-phenylpropyl, 4-phenylbutyl, α-naphthylmethyl,β-naphthylmethyl, diphenylmethyl, triphenylmethyl,α-naphthyldipheylmethyl, 9-anthrylmethyl, 4-methylbenzyl,2,4,6-trimethylbenzyl, 3,4,5-trimethylbenzyl, 4-methoxybenzyl,4-methoxyphenyldiphenylmethyl, 2-nitrobenzyl, 4-nitrobenzyl,4-chlorobenzyl, 4-bromobenzyl, 4-cyanobenzyl,4-cyanobenzyldiphenylmethyl, or bis(2-nitrophenyl)methyl groups.

The term “electron-withdrawing group” is art-recognized and describesthe ability of a substituent to attract valence electrons (e.g.,pi-electrons) from neighboring atoms, e.g., the substituent is moreelectronegative than neighboring atoms, or it draws electrons to itselfmore than a hydrogen atom would at the same position. The Hammett sigmavalue (σ) is an accepted measure of a group's electron-donating andwithdrawing ability, especially the sigma para value (σ_(p)). See, e.g.,“Advanced Organic Chemistry” by J. March, 5^(th) Ed., John Wiley & Sons,Inc., New York, pp. 368-75 (2001). The Hammett constant values aregenerally negative for electron-donating groups (σ_(p)=−0.66 for NH₂)and positive for electron-withdrawing groups (σ_(p)=0.78 for a nitrogroup), σ_(p) indicating para substitution. Exemplaryelectron-withdrawing groups include nitro, acyl (ketone), formyl(aldehyde), sulfonyl, trifluoromethyl, halogeno chloro and fluoro), andcyano groups, among others. Conversely, an “electron-donating group”designates a substituent that contributes electrons more than hydrogenwould if it occupied the same position in the molecule. Examples includeamino (including alkylamino and dialkylamino), aryl, alkoxy (includingaralkoxy), aryloxy, mercapto and alkylthio, and hydroxyl groups, amongothers.

As used herein, “alkyl” groups include saturated hydrocarbons having oneor more carbon atoms, including straight-chain alkyl groups (e.g.,methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl,decyl, etc.), cyclic alkyl groups (or “cycloalkyl” or “alicyclic” or“carbocyclic” groups) (e.g., cyclopropyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl, etc.), branched-chain alkyl groups (isopropyl,tert-butyl, sec-butyl, isobutyl, etc.), and alkyl-substituted alkylgroups (e.g., alkyl-substituted cycloalkyl groups andcycloalkyl-substituted alkyl groups). The term “aliphatic group”includes organic moieties characterized by straight or branched-chains,typically having between 1 and 22 carbon atoms. In complex structures,the chains may be branched, bridged, or cross-linked. Aliphatic groupsinclude alkyl groups, alkenyl groups, and alkynyl groups.

In certain embodiments, a straight-chain or branched-chain alkyl groupmay have 30 or fewer carbon atoms in its backbone, e.g., C₁-C₃₀ forstraight-chain or C₃-C₃₀ for branched-chain. In certain embodiments, astraight-chain or branched-chain alkyl group may have 20 or fewer carbonatoms in its backbone, e.g., C₁-C₂₀ for straight-chain or C₃-C₂₀ forbranched-chain, and more preferably 18 or fewer. Likewise, preferredcycloalkyl groups have from 4-10 carbon atoms in their ring structure,and more preferably have 4-7 carbon atoms in the ring structure. Theterm “lower alkyl” refers to alkyl groups having from 1 to 6 carbons inthe chain, and to cycloalkyl groups having from 3 to 6 carbons in thering structure.

Unless the number of carbons is otherwise specified, “lower” as in“lower aliphatic,” “lower alkyl,” “lower alkenyl,” etc. as used hereinmeans that the moiety has at least one and less than about 8 carbonatoms. In certain embodiments, a straight-chain or branched-chain loweralkyl group has 6 or fewer carbon atoms in its backbone (e.g., C₁-C₆ forstraight-chain, C₃-C₆ for branched-chain), and more preferably 4 orfewer. Likewise, preferred cycloalkyl groups have from 3-8 carbon atomsin their ring structure, and more preferably have 5 or 6 carbons in thering structure. The term “C₁-C₆” as in “C₁-C₆ alkyl” means alkyl groupscontaining 1 to 6 carbon atoms.

Moreover, unless otherwise specified the term alkyl includes both“unsubstituted alkyls” and “substituted alkyls,” the latter of whichrefers to alkyl groups having substituents replacing one or morehydrogens on one or more carbons of the hydrocarbon backbone. Suchsubstituents may include, for example, alkenyl, alkynyl, halogeno,hydroxyl, alkylcarbonyloxy, arylcarbonyloxy, alkoxycarbonyloxy, aryloxy,aryloxycarbonyloxy, carboxylate, alkylcarbonyl, arylcarbonyl,alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl,alkylthiocarbonyl, alkoxyl, phosphate, phosphonato, phosphinato, cyano,amino (including alkyl amino, dialkylamino, arylamino, diarylamino, andalkylarylamino), acylamino (including alkylcarbonylamino,arylcarbonylamino, carbamoyl and ureido), imino, sulfhydryl, alkylthio,arylthio, thiocarboxylate, sulfates, alkylsulfinyl, sulfonato,sulfamoyl, sulfonamido, nitro, trifluoromethyl, cyano, azido,heterocyclic, alkylaryl, or aromatic (including heteroaromatic) groups.

An “arylalkyl” group is an alkyl group substituted with an aryl group(e.g., phenylmethyl (i.e., benzyl)). An “alkylaryl” moiety is an arylgroup substituted with an alkyl group (e.g., p-methylphenyl (i.e.,p-tolyl)). The term “n-alkyl” means a straight-chain (i.e., unbranched)unsubstituted alkyl group. An “alkylene” group is a divalent analog ofthe corresponding alkyl group. The terms “alkenyl” and “alkynyl” referto unsaturated aliphatic groups analogous to alkyls, but which containat least one double or triple carbon-carbon bond respectively. Suitablealkenyl and alkynyl groups include groups having 2 to about 12 carbonatoms, preferably from 2 to about 6 carbon atoms.

The term “aromatic group” or “aryl group” includes unsaturated andaromatic cyclic hydrocarbons as well as unsaturated and aromaticheterocycles containing one or more rings. Aryl groups may also be fusedor bridged with alicyclic or heterocyclic rings that are not aromatic soas to form a polycycle (e.g., tetralin). An “arylene” group is adivalent analog of an aryl group. Aryl groups can also be fused orbridged with alicyclic or heterocyclic rings which are not aromatic soas to form a polycycle (e.g., tetralin).

The term “heterocyclic group” includes closed ring structures analogousto carbocyclic groups in which one or more of the carbon atoms in thering is an element other than carbon, for example, nitrogen, sulfur, oroxygen. Heterocyclic groups may be saturated or unsaturated.Additionally, heterocyclic groups (such as pyrrolyl, pyridyl,isoquinolyl, quinolyl, purinyl, and furyl) may have aromatic character,in which case they may be referred to as “heteroaryl” or“heteroaromatic” groups.

Unless otherwise stipulated, aryl and heterocyclic (includingheteroaryl) groups may also be substituted at one or more constituentatoms. Examples of heteroaromatic and heteroalicyclic groups may have 1to 3 separate or fused rings with 3 to about 8 members per ring and oneor more N, O, or S heteroatoms. In general, the term “heteroatom”includes atoms of any element other than carbon or hydrogen, preferredexamples of which include nitrogen, oxygen, sulfur, and phosphorus.Heterocyclic groups may be saturated or unsaturated or aromatic.

Examples of heterocycles include, but are not limited to, acridinyl;azocinyl; benzimidazolyl; benzofuranyl; benzothiofuranyl;benzothiophenyl; benzoxazolyl; benzthiazolyl; benztriazolyl;benztetrazolyl; benzisoxazolyl; benzisothiazolyl; benzimidazolinyl;carbazolyl; 4aH-carbazolyl; carbolinyl; chromanyl; chromenyl;cinnolinyl; decahydroquinolinyl; 2H,6H-1,5,2-dithiazinyl;dihydrofuro[2,3-b]tetrahydrofuran; furanyl; furazanyl; imidazolidinyl;imidazolinyl; imidazolyl; 1H-indazolyl; indolenyl; indolinyl;indolizinyl; indolyl; 3H-indolyl; isobenzofuranyl; isochromanyl;isoindazolyl; isoindolinyl; isoindolyl; isoquinolinyl; isothiazolyl;isoxazolyl; methylenedioxyphenyl; morpholinyl; naphthyridinyl;octahydroisoquinolinyl; oxadiazolyl; 1,2,3-oxadiazolyl;1,2,4-oxadiazolyl; 1,2,5-oxadiazolyl; 1,3,4-oxadiazolyl; oxazolidinyl;oxazolyl; oxazolidinyl; pyrimidinyl; phenanthridinyl; phenanthrolinyl;phenazinyl; phenothiazinyl; phenoxathiinyl; phenoxazinyl; phthalazinyl;piperazinyl; piperidinyl; piperidonyl; 4-piperidonyl; piperonyl;pteridinyl; purinyl; pyranyl; pyrazinyl; pyrazolidinyl; pyrazolinyl;pyrazolyl; pyridazinyl; pyridooxazole; pyridoimidazole; pyridothiazole;pyridinyl; pyridyl; pyrimidinyl; pyrrolidinyl; pyrrolinyl; 2H-pyrrolyl;pyrrolyl; quinazolinyl; quinolinyl; 4H-quinolizinyl; quinoxalinyl;quinuclidinyl; tetrahydrofuranyl; tetrahydroisoquinolinyl;tetrahydroquinolinyl; tetrazolyl; 6H-1,2,5-thiadiazinyl;1,2,3-thiadiazolyl; 1,2,4-thiadiazolyl; 1,2,5-thiadiazolyl;1,3,4-thiadiazolyl; thianthrenyl; thiazolyl; thienyl; thienothiazolyl;thienooxazolyl; thienoimidazolyl; thiophenyl; triazinyl;1,2,3-triazolyl; 1,2,4-triazolyl; 1,2,5-triazolyl; 1,3,4-triazolyl; andxanthenyl. Preferred heterocycles include, but are not limited to,pyridinyl; furanyl; thienyl; pyrrolyl; pyrazolyl; pyrrolidinyl;imidazolyl; indolyl; benzimidazolyl; 1H-indazolyl; oxazolidinyl;benzotriazolyl; benzisoxazolyl; oxindolyl; benzoxazolinyl; and isatinoylgroups. Also included are fused ring and spiro compounds containing, forexample, the above heterocycles.

A common hydrocarbon aryl group is a phenyl group having one ring.Two-ring hydrocarbon aryl groups include naphthyl, indenyl,benzocyclooctenyl, benzocycloheptenyl, pentalenyl, and azulenyl groups,as well as the partially hydrogenated analogs thereof such as indanyland tetrahydronaphthyl. Exemplary three-ring hydrocarbon aryl groupsinclude acephthylenyl, fluorenyl, phenalenyl, phenanthrenyl, andanthracenyl groups.

Aryl groups also include heteromonocyclic aryl groups, i.e., single-ringheteroaryl groups, such as thienyl, furyl, pyranyl, pyrrolyl,imidazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, andpyridazinyl groups; and oxidized analogs thereof such as pyridonyl,oxazolonyl, pyrazolonyl, isoxazolonyl, and thiazolonyl groups. Thecorresponding hydrogenated (i.e., non-aromatic) heteromonocylic groupsinclude pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl,pyrazolidinyl, pyrazolinyl, piperidyl and piperidino, piperazinyl, andmorpholino and morpholinyl groups.

Aryl groups also include fused two-ring heteroaryls such as indolyl,isoindolyl, indolizinyl, indazolyl, quinolinyl, isoquinolinyl,phthalazinyl, quinoxalinyl, quinazolinyl, cinnolinyl, chromenyl,isochromenyl, benzothienyl, benzimidazolyl, benzothiazolyl, purinyl,quinolizinyl, isoquinolonyl, quinolonyl, naphthyridinyl, and pteridinylgroups, as well as the partially hydrogenated analogs such as chromanyl,isochromanyl, indolinyl, isoindolinyl, and tetrahydroindolyl groups.Aryl groups also include fused three-ring groups such as phenoxathiinyl,carbazolyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl,phenazinyl, phenothiazinyl, phenoxazinyl, and dibenzofuranyl groups.

Some typical aryl groups include substituted or unsubstituted 5- and6-membered single-ring groups. In another aspect, each Ar group may beselected from the group consisting of substituted or unsubstitutedphenyl, pyrrolyl, furyl, thienyl, thiazolyl, isothiaozolyl, imidazolyl,triazolyl, tetrazolyl, pyrazolyl, oxazolyl, isooxazolyl, pyridinyl,pyrazinyl, pyridazinyl, and pyrimidinyl groups. Further examples includesubstituted or unsubstituted phenyl, 1-naphthyl, 2-naphthyl, biphenyl,1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl,4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl,3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl,5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl,3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl,purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl groups.

The term “amine” or “amino,” as used herein, refers to an unsubstitutedor substituted moiety of the formula —NR^(a)R^(b), in which R^(a) andR^(b) are each independently hydrogen, alkyl, aryl, or heterocyclyl, orR^(a) and R^(b), taken together with the nitrogen atom to which they areattached, form a cyclic moiety having from 3 to 8 atoms in the ring.Thus, the term amino includes cyclic amino moieties such as piperidinylor pyrrolidinyl groups, unless otherwise stated. Thus, the term“alkylamino” as used herein means an alkyl group having an amino groupattached thereto. Suitable alkylamino groups include groups having 1 toabout 12 carbon atoms, preferably from 1 to about 6 carbon atoms. Theterm amino includes compounds or moieties in which a nitrogen atom iscovalently bonded to at least one carbon or heteroatom. The term“dialkylamino” includes groups wherein the nitrogen atom is bound to atleast two alkyl groups. The term “arylamino” and “diarylamino” includegroups wherein the nitrogen is bound to at least one or two aryl groups,respectively. The term “alkylarylamino” refers to an amino group whichis bound to at least one alkyl group and at least one aryl group. Theterm “alkaminoalkyl” refers to an alkyl, alkenyl, or alkynyl groupsubstituted with an alkylamino group. The term “amide” or“aminocarbonyl” includes compounds or moieties which contain a nitrogenatom which is bound to the carbon of a carbonyl or a thiocarbonyl group.

The term “alkylthio” refers to an alkyl group, having a sulfhydryl groupattached thereto. Suitable alkylthio groups include groups having 1 toabout 12 carbon atoms, preferably from 1 to about 6 carbon atoms.

The term “alkylcarboxyl” as used herein means an alkyl group having acarboxyl group attached thereto.

The term “alkoxy” as used herein means an alkyl group having an oxygenatom attached thereto. Representative alkoxy groups include groupshaving 1 to about 12 carbon atoms, preferably 1 to about 6 carbon atoms,e.g., methoxy, ethoxy, propoxy, tert-butoxy and the like. Examples ofalkoxy groups include methoxy, ethoxy, isopropyloxy, propoxy, butoxy,and pentoxy groups. The alkoxy groups can be substituted with groupssuch as alkenyl, alkynyl, halogen, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moieties. Examples of halogen substituted alkoxygroups include, but are not limited to, fluoromethoxy, difluoromethoxy,trifluoromethoxy, chloromethoxy, dichloromethoxy, trichloromethoxy,etc., as well as perhalogenated alkyloxy groups.

The term “acylamino” includes moieties wherein an amino moiety is bondedto an acyl group. For example, the acylamino group includesalkylcarbonylamino, arylcarbonylamino, carbamoyl and ureido groups.

The terms “alkoxyalkyl”, “alkylaminoalkyl” and “thioalkoxyalkyl” includealkyl groups, as described above, which further include oxygen, nitrogenor sulfur atoms replacing one or more carbons of the hydrocarbonbackbone.

The term “carbonyl” or “carboxy” includes compounds and moieties whichcontain a carbon connected with a double bond to an oxygen atom.Examples of moieties which contain a carbonyl include aldehydes,ketones, carboxylic acids, amides, esters, anhydrides, etc.

The term “ether” or “ethereal” includes compounds or moieties whichcontain an oxygen bonded to two carbon atoms. For example, an ether orethereal group includes “alkoxyalkyl” which refers to an alkyl, alkenyl,or alkynyl group substituted with an alkoxy group.

A “sulfonate” group is a —SO₃H or —SO₃ ⁻X⁺ group bonded to a carbonatom, where X⁺ is a cationic counter ion group. Similarly, a “sulfonicacid” compound has a —SO₃H or —SO₃ ⁻X⁺ group bonded to a carbon atom,where X+ is a cationic group. A “sulfate” as used herein is a —OSO₃H or—OSO₃ ⁻X⁺ group bonded to a carbon atom, and a “sulfuric acid” compoundhas a —SO₃H or −OSO₃ ⁻X⁺ group bonded to a carbon atom, where X⁺ is acationic group. According to the invention, a suitable cationic groupmay be a hydrogen atom. In certain cases, the cationic group mayactually be another group on the therapeutic compound that is positivelycharged at physiological pH, for example an amino group.

A “counter ion” is required to maintain electroneutrality. Examples ofanionic counter ions include halide, triflate, sulfate, nitrate,hydroxide, carbonate, bicarbonate, acetate, phosphate, oxalate, cyanide,alkylcarboxylate, N-hydroxysuccinimide, N-hydroxybenzotriazole,alkoxide, thioalkoxide, alkane sulfonyloxy, halogenated alkanesulfonyloxy, arylsulfonyloxy, bisulfate, oxalate, valerate, oleate,palmitate, stearate, laurate, borate, benzoate, lactate, citrate,maleate, fumarate, succinate, tartrate, naphthylate mesylate,glucoheptonate, or lactobionate. Compounds containing a cationic groupcovalently bonded to an anionic group may be referred to as an “internalsalt.”

The term “nitro” means —NO₂; the term “halogen” or “halogeno” or “halo”designates —F, —Cl, —Br or —I; the term “thiol,” “thio,” or “mercapto”means SH; and the term “hydroxyl” or “hydroxy” means —OH.

The term “acyl” refers to a carbonyl group that is attached through itscarbon atom to a hydrogen (i.e., a formyl), an aliphatic group (e.g.,acetyl), an aromatic group (e.g., benzoyl), and the like. The term“substituted acyl” includes acyl groups where one or more of thehydrogen atoms on one or more carbon atoms are replaced by, for example,an alkyl group, alkynyl group, halogen, hydroxyl, alkylcarbonyloxy,arylcarbonyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, carboxylate,alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, aminocarbonyl,alkylaminocarbonyl, dialkylaminocarbonyl, alkylthiocarbonyl, alkoxyl,phosphate, phosphonato, phosphinato, cyano, amino (including alkylamino, dialkylamino, arylamino, diarylamino, and alkylarylamino),acylamino (including alkylcarbonylamino, arylcarbonylamino, carbamoyland ureido), imino, sulfhydryl, alkylthio, arylthio, thiocarboxylate,sulfates, alkylsulfinyl, sulfonato, sulfamoyl, sulfonamido, nitro,trifluoromethyl, cyano, azido, heterocyclyl, alkylaryl, or an aromaticor heteroaromatic moiety.

Unless otherwise specified, the chemical moieties of the compounds ofthe invention, including those groups discussed above, may be“substituted or unsubstituted.” In some embodiments, the term“substituted” means that the moiety has substituents placed on themoiety other than hydrogen (i.e., in most cases, replacing a hydrogen),which allow the molecule to perform its intended function. Examples ofsubstituents include moieties selected from straight or branched alkyl(preferably C₁-C₅), cycloalkyl (preferably C₃-C₈), alkoxy (preferablyC₁-C₆), thioalkyl (preferably C₁-C₆), alkenyl (preferably C₂-C₆),alkynyl (preferably C₂-C₆), heterocyclic, carbocyclic, aryl (e.g.,phenyl), aryloxy (e.g., phenoxy), aralkyl (e.g., benzyl), aryloxyalkyl(e.g., phenyloxyalkyl), arylacetamidoyl, alkylaryl, heteroaralkyl,alkylcarbonyl and arylcarbonyl or other such acyl group,heteroarylcarbonyl, and heteroaryl groups, as well as (CR′R″)₀₋₃NR′R″(e.g., —NH₂), (CR′R″)₀₋₃CN (e.g., —CN), —NO₂, halogen (e.g., —F, —Cl,—Br, or —I), (CR′R″)₀₋₃C(halogen)₃ (e.g., —CF₃), (CR′R″)₀₋₃CH(halogen)₂,(CR′R″)₀₋₃CH₂(halogen), (CR′R″)₀₋₃CONR′R″, (CR′R″)₀₋₃(CNH)NR′R″,(CR′R″)₀₋₃S(O)₁₋₂NR′R″, (CR′R″)₀₋₃CHO, (CR′R″)₀₋₃O(CR′R″)₀₋₃H,(CR′R″)₀₋₃S(O)₀₋₃R′ (e.g., —SO₃H), (CR′R″)₀₋₃O(CR′R″)₀₋₃H (e.g.,—CH₂OCH₃ and —OCH₃), (CR′R″)₀₋₃S(CR′R″)₀₋₃H (e.g., —SH and —SCH₃),(CR′R″)₀₋₃OH (e.g., —OH), (CR′R″)₀₋₃COR′, (CR′R″)₀₋₃(substituted orunsubstituted phenyl), (CR′R″)₀₋₃(C₃-C₈ cycloalkyl), (CR′R″)₀₋₃CO₂R′(e.g., —CO₂H), and (CR′R″)₀₋₃OR′ groups, wherein R′ and R″ are eachindependently hydrogen, a C₁-C₅ alkyl, C₂-C₅ alkenyl, C₂-C₅ alkynyl, oraryl group; or the side chain of any naturally occurring amino acid.

In another embodiment, a substituent may be selected from straight orbranched alkyl (preferably C₁-C₅), cycloalkyl (preferably C₃-C₈), alkoxy(preferably C₁-C₆), thioalkyl (preferably C₁-C₆), alkenyl (preferablyC₂-C₆), alkynyl (preferably C₂-C₆), heterocyclic, carbocyclic, aryl(e.g., phenyl), aryloxy (e.g., phenoxy), aralkyl (e.g., benzyl),aryloxyalkyl (e.g., phenyloxyalkyl), arylacetamidoyl, alkylaryl,heteroaralkyl, alkylcarbonyl and arylcarbonyl or other such acyl group,heteroarylcarbonyl, or heteroaryl group, (CR′R″)₀₋₁₀NR′R″ (e.g., —NH₂),(CR′R″)₀₋₁₀CN (e.g., —CN), NO₂, halogen (e.g., F, Cl, Br, or I),(CR′R″)₀₋₁₀C(halogen)₃ (e.g., —CF₃), (CR′R″)₀₋₁₀CH(halogen)₂,(CR′R″)₀₋₁₀CH₂(halogen), (CR′R″)₀₋₁₀CONR′R″, (CR′R″)₀₋₁₀(CNH)NR′R″,(CR′R″)₀₋₁₀S(O)₁₋₂NR′R″, (CR′R″)₀₋₁₀CHO, (CR′R″)₀₋₁₀O(CR′R″)₀₋₁₀H,(CR′R″)₀₋₁₀S(O)₀₋₃R′ (e.g., —SO₃H), (CR′R″)₀₋₁₀O(CR′R″)₀₋₁₀H (e.g.,—CH₂OCH₃ and —OCH₃), (CR′R″)₀₋₁₀S(CR′R″)₀₋₃H (e.g., —SH and —SCH₃),(CR′R″)₀₋₁₀OH (e.g., —OH), (CR′R″)₀₋₁₀OOR′, (CR′R″)₀₋₁₀ (substituted orunsubstituted phenyl), (CR′R″)₀₋₁₀(C₃-C₈ cycloalkyl), (CR′R″)₀₋₁₀CO₂R′(e.g., —CO₂H), or (CR′R″)₀₋₁₀₀R′ group, or the side chain of anynaturally occurring amino acid; wherein R′ and R″ are each independentlyhydrogen, a C₁-C₅ alkyl, C₂-C₅ alkenyl, C₂-C₅ alkynyl, or aryl group, orR′ and R″ taken together are a benzylidene group or a —(CH₂)₂—O—(CH₂)₂—group.

It will be understood that “substitution” or “substituted with” includesthe implicit proviso that such substitution is in accordance with thepermitted valence of the substituted atom and the substituent, and thatthe substitution results in a stable compound, e.g., which does notspontaneously undergo transformation such as by rearrangement,cyclization, elimination, etc. As used herein, the term “substituted” ismeant to include all permissible substituents of organic compounds. In abroad aspect, the permissible substituents include acyclic and cyclic,branched and unbranched, carbocyclic and heterocyclic, aromatic andnonaromatic substituents of organic compounds. The permissiblesubstituents can be one or more.

In some embodiments, a “substituent” may be selected from the groupconsisting of, for example, halogeno, trifluoromethyl, nitro, cyano,C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, C₁-C₆ alkylcarbonyloxy,arylcarbonyloxy, C₁-C₆ alkoxycarbonyloxy, aryloxycarbonyloxy, C₁-C₆alkylcarbonyl, C₁-C₆ alkoxycarbonyl, C₁-C₆ alkoxy, C₁-C₆ alkylthio,arylthio, heterocyclyl, aralkyl, and aryl (including heteroaryl) groups.

In one embodiment, the invention pertains to compounds of Formula I:

wherein:

R¹ is a substituted or unsubstituted cycloalkyl, heterocyclic, aryl,arylcycloalkyl, bicyclic or tricyclic ring, a bicyclic or tricyclicfused ring group, or a substituted or unsubstituted C₂-C₁₀ alkyl group;

R² is selected from a group consisting of hydrogen, alkyl,mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl,triazolyl, imidazolyl, benzothiazolyl, and benzoimidazolyl;

Y is SO₃ ⁻X⁺, OSO₃ ⁻X⁺, or SSO₃ ⁻X⁺;

X⁺ is hydrogen, a cationic group, or ester-forming group; and

each of L¹ and L² is independently a substituted or unsubstituted C₁-C₅alkyl group or absent, or a pharmaceutically acceptable salt thereof,provided that when R₁ is alkyl, L¹ is absent.

In a further embodiment, the invention pertains to compounds of FormulaII:

wherein:

R¹ is a substituted or unsubstituted cyclic, bicyclic, tricyclic, orbenzoheterocyclic group or a substituted or unsubstituted C₂-C₁₀ alkylgroup;

R² is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl,aryl, arylalkyl, thiazolyl, triazolyl, imidazolyl, benzothiazolyl,benzoimidazolyl, or linked to R¹ to form a heterocycle;

Y is SO₃ ⁻X⁺, OSO₃ ⁻X⁺, or SSO₃ ⁻X⁺;

X⁺ is hydrogen, a cationic group, or an ester forming moiety;

m is 0 or 1;

n is 1, 2, 3, or 4;

L is substituted or unsubstituted C₁-C₃ alkyl group or absent, or apharmaceutically acceptable salt thereof, provided that when R¹ isalkyl, L is absent.

In a further embodiment, R² is hydrogen. In another further embodiment,R¹ is straight chain alkyl, for example, ethyl, n-pentyl, n-heptyl, orn-octyl. In another embodiment, R¹ is t-butyl. In yet another alternateembodiment, R¹ is C₇-C₁₀ bicycloalkyl or tricycloalkyl, such as, forexample, tricyclo[3.3.1.0^(3,7)]decyl (or adamantyl),bicyclo[2.1.2]heptyl, or indolyl. In another alternate embodiment, R¹ istetrahydronaphthyl.

In one embodiment, L² is —(CH₂)₃—. In another further embodiment, L² is—(CH₂)₄— or —(CH₂)₅—. In yet another further embodiment, L₂ is —(CH₂)₂—.In yet another further embodiment, L² is substituted alkyl, e.g.,—CH₂—(CHOH)—CH₂—.

In another embodiment, L′ is CH₂CH₂ or absent.

In a further embodiment, R¹ is branched alkyl, e.g., t-butyl. In anotherembodiment, R¹ is adamanyl. In another embodiment, R¹ is cyclic alkyl,e.g., cyclopropyl, cyclohexyl, cycloheptyl, cyclo-octyl, etc. Thecycloalkyl moieties may be substituted further, e.g., with additionalalkyl groups or other groups which allow the molecule to perform itsintended function. In another embodiment, R¹ is alkyl substituted with apropargyl moiety (e.g., HC≡C—). In another embodiment, R¹ is cyclohexylsubstituted withone or more methyl or propargyl groups.

In other embodiments, L¹ is a C₁-C₂ alkyl linker group (e.g., —CH(CH₃)—or —(CH₂)₂—. In a further embodiment, R¹ is phenyl. In certainembodiments, R¹ is substituted with a methoxy group. In otherembodiments, L¹ is C₃, e.g., —(CH₂)₃— or C(CH₃)₂—. In certainembodiments, L¹ is substituted, e.g., with an alkoxy, carboxylate(—COOH), benzyl, amido (—C═O—NH—), or ester (C═O—C—O) group. In certainembodiment, the ester group is a methyl, ethyl, propyl, butyl,cyclohexyl, or benzyl ester. In other embodiments, the ester group maybe propenyl. In other embodiments, L¹ is substituted with a carboxylategroup. In a further embodiment, R¹ is substituted with a substitutedamido group, wherein the amido group is substituted with an alkyl, e.g.,methyl, ethyl, propyl, butyl, pentyl, or hexyl group. In anotherembodiment, the alkyl R¹ group is a substituted with a —C═O—NH—OH,C═O—NH₂, or an amido group. In certain embodiments, the amido group issubstituted with an alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl,hexyl, cyclohexyl, etc.), a benzyl or an aryl group. In anotherembodiment, the amido group is substituted with a —CH(CH₂)₂ group. R¹itself may be substituted with a phenyl or may be branched or straightchain alkyl. In certain embodiments, R¹ may also be substituted with athioether moiety. Examples of thioethers include S-Me, S-Et, etc. Incertain embodiments, the alkyl R¹ moiety is substituted with both anaryl or a thioether moiety and an amido moiety. In other embodiments,the alkyl R¹ moiety may be substituted with both a thioether and acarboxylate moiety. In other embodiments, alkyl R¹ groups aresubstituted with hydroxyl. R¹ groups, e.g., alkyl R¹ groups, may also besubstituted with both thioether and hydroxyl groups. In otherembodiments, R¹ groups, e.g., alkyl R¹ groups are substituted with cyanogroups. Examples of R¹ groups including —CN moieties include —C(CH₃)₂CN,cyclohexyl substituted with one or more cyano groups, etc.

In other embodiments, alkyl R¹ groups are substituted with aryl groups.The aryl groups may be substituted phenyl, for example. The substitutedphenyl may be substituted with one or more substituents such as hydroxy,cyano and alkoxy. In other embodiments, alkyl R¹ groups are substitutedwith tetrazolyl or substituted or unsubstituted benzyl.

In a further embodiment, L^(I) is —C(CH₃)₂—(CH₂)—. In anotherembodiment, L^(I) is —(C(CH₃)₂—CHOH—. In yet another embodiment, L^(I)is —(C(CH₃)₂CH(OMe)—. In another embodiment, R¹ is substituted orunsubstituted phenyl. In a further embodiment, R¹ is para-substitutedphenyl. Examples of substitutuents include but are not limited tofluorine, chlorine, bromine, iodine, methyl, t-butyl, alkoxy, methoxy,etc. In other embodiment, R¹ is substituted at the meta position.Examples of substituents include methoxy, chloro, methyl, t-butyl,fluoro, alkyl, alkoxy, iodo, trifluoroalkyl, methoxy, etc. In anotherembodiment, R¹ is phenyl substituted in the ortho position, with similarsubstituents. In another embodiment, L¹ comprises a cycloalkyl moiety,e.g., cyclopentyl. In another embodiment, L¹ comprises an alkyenyl groupand, optionally, a substituted aryl group, with substitutents similar tothose described about.

In certain embodiments, R¹ is cyclopropyl or cyclohexyl. In certainembodiments, the cyclopropyl or cyclohexyl group is substituted with anether group or an alkyl group. In certain further embodiments, the ethergroup is a benzyl ether group.

In another embodiment, wherein R¹ is alkyl, it is substituted withgroups such as phenyl, or hydroxy.

In other embodiments, the compound of the invention is selected from thegroup consisting of:

and pharmaceutically acceptable salts, esters, and prodrugs thereof.

In another embodiment, the invention pertains to compounds of FormulaIII:

wherein:

A is nitrogen or oxygen;

R¹¹ is hydrogen, salt-forming cation, ester forming group, —(CH₂)_(x)-Q,or when A is nitrogen, A and R¹¹ taken together may be the residue of anatural or unnatural amino acid or a salt or ester thereof;

Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, orbenzoimidazolyl;

x is 0, 1, 2, 3, or 4;

n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

R³, R^(3a), R⁴, R^(4a), R⁵, R^(5a), R⁶, R^(6a), R⁷ and R^(7a) are eachindependently hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl,cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, cyano,halogen, amino, tetrazolyl, or two R groups on adjacent ring atoms takentogether with the ring atoms form a double bond, provided that one ofR³,R^(3a), R⁴, R^(4a), R⁵, R^(5a), R⁶, R^(6a), R⁷ and R^(7a) is a moietyof the Formula

wherein:

m is 0, 1, 2, 3, or 4;

R^(A′), R^(B′), R^(C′), R^(D′), and R^(E′) re independently selectedfrom a group of hydrogen, halogen, hydroxyl, alkyl, alkoxyl, halogenatedalkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, cyano,thiazolyl, triazolyl, imidazolyl, tetrazolyl, benzothiazolyl, andbenzoimidazolyl; and pharmaceutically acceptable salts and estersthereof, provided that said compound is not3-(4-phenyl-1,2,3,6-tetrahydro-1-pyridyl)-1-propanesulfonic acid.

In a further embodiment, n is 2, 3 or 4.

In another embodiment, R¹¹ is a salt-forming cation. Examples of saltforming cations include pharmaceutically acceptable salts describedherein as well as lithium, sodium, potassium, magnesium, calcium,barium, zinc, iron, and ammonium. In another embodiment, R¹¹ is anester-forming group. An ester-forming group includes groups which whenbound form an ester. Examples of such groups include substituted orunsubstituted alkyl, aryl, alkenyl, alkynyl, or cycloalkyl. In anotherembodiment, A is oxygen.

In another embodiment, R³ and R⁴ are taken together with the carbonatoms to which they are attached to form a double bond. In anotherembodiment, R^(A′), R^(B′), R^(C′), R^(D′), and R^(E′) are eachhydrogen. R^(A′), R^(B′), R^(D′), and R^(E′) are each hydrogen andR^(C′) is a halogen, such as fluorine, chlorine, iodine, or bromine.

In another embodiment, R³ or R^(5a) is a moiety of Formula IIIa.

In another embodiment, R⁴, R⁵, R⁶, and R⁷ are each hydrogen. In anotherfurther embodiment, R^(4a), R^(5a), R^(6a), and R^(7a) are eachhydrogen.

In another, R^(3a) is hydroxyl, cyano, acyl, or hydroxyl.

In another further embodiment, R¹¹ and A taken together are a natural orunnatural amino acid residue or a pharmaceutically acceptable salt orester thereof. Examples of amino acid residues include esters and saltsof phenylalanine and leucine.

In another embodiment, m is 0, 1, or 3.

Examples of compounds of Formula III include, but are not limited to:

and pharmaceutically acceptable salts, esters, and prodrugs thereof.

In another embodiment, the invention pertains to compounds of FormulaIV:

wherein:

A is nitrogen or oxygen;

R¹¹ is hydrogen, salt-forming cation, ester forming group, —(CH₂)_(x)-Q,or when A is nitrogen, A and R¹¹ taken together may be the residue of anatural or unnatural amino acid or a salt or ester thereof;

Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, orbenzoimidazolyl;

x is 0, 1, 2, 3, or 4;

n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

R⁴, R^(4a), R⁵, R^(5a), R⁶, R^(6a), R⁷, and R^(7a) are eachindependently hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl,cycloalkyl, aryl, alkylcarbonyl, arylcarbonyl, alkoxycarbonyl, cyano,halogen, amino, tetrazolyl, R⁴ and R⁵ taken together, with the ringatoms they are attached to, form a double bond, or R⁶ and R⁷ takentogether, with the ring atoms they are attached to, form a double bond;

m is 0, 1, 2, 3, or 4;

R⁸, R⁹, R¹⁹, R¹¹, and R¹² are independently selected from a group ofhydrogen, halogen, hydroxyl, alkyl, alkoxyl, halogenated alkyl,mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, cyano, thiazolyl,triazolyl, imidazolyl, tetrazolyl, benzothiazolyl, and benzoimidazolyl,and pharmaceutically acceptable salts and esters thereof.

In another embodiment, R¹¹ is a salt-forming cation. Examples of saltforming cations include pharmaceutically acceptable salts describedherein as well as lithium, sodium, potassium, magnesium, calcium,barium, zinc, iron, and ammonium. In another embodiment, R¹¹ is anester-forming group. An ester-forming group includes groups which whenbound form an ester. Examples of such groups include substituted orunsubstituted alkyl, aryl, alkenyl, alkynyl, or cycloalkyl. In anotherembodiment, A is oxygen.

In another embodiment, m is 0 or 1. In another further embodiment, n is2, 3, or 4. In another further embodiment, R⁴, R⁵, R⁶ and R⁷ are eachhydrogen. R^(4a), R^(5a), R^(6a), and R^(7a) also may be hydrogen.Examples of R⁸, R⁹, R¹⁰, R¹¹, and R¹² include hydrogen. In anotherembodiment R⁸, R⁹, R¹¹, R¹² are each hydrogen, and R¹⁰ is a halogen,(e.g., fluorine, chlorine, bromine, or iodine), nitro, or alkyl (e.g.,methyl, ethyl, butyl).

In another embodiment, A-R¹¹ may be the residue of an amino acid, e.g.,a phenylalanine residue. In another embodiment, R⁹, R¹⁰, R¹¹ and R¹² areeach hydrogen, and R⁸ is not hydrogen, e.g., halogen, e.g., fluorine,bromine, chlorine, or iodine.

In another embodiment, the compound is:

and pharmaceutically acceptable salts, esters, and prodrugs thereof.

In another embodiment, the invention pertains to compounds of Formula V:

wherein:

A is nitrogen or oxygen;

R¹¹ is hydrogen, salt-forming cation, ester forming group, —(CH₂)_(x)-Q,or when A is nitrogen, A and R¹¹ taken together may be the residue of anatural or unnatural amino acid or a salt or ester thereof;

Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, orbenzoimidazolyl;

x is 0, 1, 2, 3, or 4;

n is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

aa is a natural or unnatural amino acid residue;

m is 0, 1, 2, or 3;

R¹⁴ is hydrogen or protecting group;

R¹⁵ is hydrogen, alkyl or aryl, and pharmaceutically acceptable salts,esters and prodrugs thereof.

In another embodiment, R¹¹ is a salt-forming cation. Examples of saltforming cations include pharmaceutically acceptable salts describedherein as well as lithium, sodium, potassium, magnesium, calcium,barium, zinc, iron, and ammonium. In another embodiment, R¹¹ is anester-forming group. An ester-forming group includes groups which whenbound form an ester. Examples of such groups include substituted orunsubstituted alkyl, aryl, alkenyl, alkynyl, or cycloalkyl. In anotherembodiment, A is oxygen.

In an embodiment, n is 2, 3 or 4. In certain embodiments, m is 0. Incertain embodiments, A-R¹¹ is a residue of a natural amino acid, or asalt or ester thereof. Examples of amino acid residues, include, but arenot limited to, leucine or phenylalanine residues, and pharmaceuticallyacceptable salts and esters thereof. Examples of possible esters includemethyl, ethyl, and t-butyl.

In another embodiment, m is 1. Examples of aa include natural andunnatural amino acid residues such as phenylalanine, glycine, andleucine.

In another embodiment, (aa)_(m) is a residue of phe-phe, or an esterthereof.

In certain embodiments, R¹⁵ is hydrogen or substituted alkyl, e.g.,arylalkyl.

The term “unnatural amino acid” refers to any derivative of a naturalamino acid including D forms, and α- and β-amino acid derivatives. Theterms “unnatural aminoacid” and “non-natural amino acid” are usedinterchangably herein and are meant to include the same moieties. It isnoted that certain amino acids, e.g., hydroxyproline, that areclassified as a non-natural amino acid herein, may be found in naturewithin a certain organism or a particular protein. Amino acids with manydifferent protecting groups appropriate for immediate use in the solidphase synthesis of peptides are commercially available. In addition tothe twenty most common naturally occurring amino acids, the followingexamples of non-natural amino acids and amino acid derivatives may beused according to the invention (common abbreviations in parentheses):β-alanine (β-ALA), γ-aminobutyric acid (GABA), 2-aminobutyric acid(2-Abu), α,β-dehydro-2-aminobutyric acid (8-AU),1-aminocyclopropane-1-carboxylic acid (ACPC), aminoisobutyric acid(Aib), 2-amino-thiazoline-4-carboxylic acid, 5-aminovaleric acid(5-Ava), 6-aminohexanoic acid (6-Ahx), 8-aminooctanoic acid (8-Aoc),11-aminoundecanoic acid (11-Aun), 12-aminododecanoic acid (12-Ado),2-aminobenzoic acid (2-Abz), 3-aminobenzoic acid (3-Abz), 4-aminobenzoicacid(4-Abz), 4-amino-3-hydroxy-6-methylheptanoic acid (Statine, Sta),aminooxyacetic acid (Aoa), 2-aminotetraline-2-carboxylic acid (ATC),4-amino-5-cyclohexyl-3-hydroxypentanoic acid (ACHPA),para-aminophenylalanine (4-NH₂-Phe), biphenylalanine (Bip),para-bromophenylalanine (4-Br-Phe), ortho-chlorophenylalanine](2-Cl-Phe), meta-chlorophenylalanine (3-C1-Phe),para-chlorophenylalanine (4-Cl-Phe), meta-chlorotyrosine (3-C1-Tyr),para-benzoylphenylalanine (Bpa), tert-butylglycine (TLG),cyclohexylalanine (Cha), cyclohexylglycine (Chg), 2,3-diaminopropionicacid (Dpr), 2,4-diaminobutyric acid (Dbu), 3,4-dichlorophenylalanine(3,4-C₁₂-Phe), 3,4-difluororphenylalanine (3,4-F₂-Phe),3,5-diiodotyrosine (3,5-I₂-Tyr), ortho-fluorophenylalanine (2-F-Phe),meta-fluorophenylalanine (3-F-Phe), para-fluorophenylalanine (4-F-Phe),meta-fluorotyrosine (3-F-Tyr), homoserine (Hse), homophenylalanine(Hfe), homotyrosine (Htyr), 5-hydroxytryptophan (5-OH-Trp),hydroxyproline (Hyp), para-iodophenylalanine (4-I-Phe), 3-iodotyrosine(34-Tyr), indoline-2-carboxylic acid (Idc), isonipecotic acid (Inp),meta-methyltyrosine (3-Me-Tyr), 1-naphthylalanine (1-Nal),2-naphthylalanine (2-Nal), para-nitrophenylalanine (4-NO₂-Phe),3-nitrotyrosine (3-NO₂-Tyr), norleucine (Nle), norvaline (Nva),ornithine (Om), ortho-phosphotyrosine (H₂PO₃-Tyr),octahydroindole-2-carboxylic acid (Oic), penicillamine (Pen),pentafluorophenylalanine (F₅-Phe), phenylglycine (Phg), pipecolic acid(Pip), propargylglycine (Pra), pyroglutamic acid (PGLU), sarcosine(Sar), tetrahydroisoquinoline-3-carboxylic acid (Tic), thienylalanine,and thiazolidine-4-carboxylic acid (thioproline, Th). Additionally,N-alkylated amino acids may be used, as well as amino acids havingamine-containing side chains (such as Lys and Orn) in which the aminehas been acylated or alkylated.

Examples of compounds of the invention include, but are not limited to.

and pharmaceutically acceptable salts, esters, and prodrugs thereof.

In another embodiment, the invention pertains, at least in part, tocompounds of Formula VI:

wherein:

n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

A is oxygen or nitrogen;

R¹¹ is hydrogen, salt-forming cation, ester forming group, —(CH₂)_(x)-Q,or when A is nitrogen, A and R¹¹ taken together may be the residue of anatural or unnatural amino acid or a salt or ester thereof;

Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, orbenzoimidazolyl;

x is 0, 1, 2, 3, or 4;

R¹⁹ is hydrogen, alkyl or aryl;

Y¹ is oxygen, sulfur, or nitrogen;

Y² is carbon, nitrogen, or oxygen;

R²⁰ is hydrogen, alkyl, amino, mercaptoalkyl, alkenyl, alkynyl,cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl,imidazolyl, benzothiazolyl, or benzoimidazolyl;

R²¹ is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl,aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl, imidazolyl,benzothiazolyl, benzoimidazolyl, or absent if Y² is oxygen;

R²² is hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl,aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl, imidazolyl,benzothiazolyl, benzoimidazolyl; or R²² is hydrogen, hydroxyl, alkoxy oraryloxy if Y¹ is nitrogen; or R²² is absent if Y¹ is oxygen or sulfur;or R²² and R²¹ may be linked to form a cyclic moiety if Y¹ is nitrogen;

R²³ is hydrogen, alkyl, amino, mercaptoalkyl, alkenyl, alkynyl,cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl, tetrazolyl,imidazolyl, benzothiazolyl, or benzoimidazolyl, or absent if Y² isnitrogen or oxygen;

or pharmaceutically acceptable salts thereof.

In another embodiment, R¹¹ is a salt-forming cation. Examples of saltforming cations include pharmaceutically acceptable salts describedherein as well as lithium, sodium, potassium, magnesium, calcium,barium, zinc, iron, and ammonium. In a further embodiment, the salt is asodium salt. In a further, embodiment, A is oxygen.

In another embodiment, Y¹ is oxygen or sulfur, and R²² is absent.

In another embodiment, Y² is oxygen and R²¹ is absent. Examples of R²⁰include benzyl, aryl (e.g., phenyl), alkyl, cycloalkyl (e.g.,adamantyl), etc. In other embodiment, Y² is nitrogen and R²¹ ishydrogen. In other embodiment, R²¹ is benzyl. In another furtherembodiment, R²⁰ and R²¹ are linked to form a pyridyl ring. In anotherembodiment, Y¹ is sulfur.

Examples of compounds of the invention, include

and pharmaceutically acceptable salts, esters, and prodrugs thereof.

In another embodiment, the invention pertains to compounds of FormulaVII:

wherein:

n is 2, 3, or 4;

A is oxygen or nitrogen;

R¹¹ is hydrogen, salt-forming cation, ester forming group, —(CH₂)_(x)-Q,or when A is nitrogen, A and R¹¹ taken together may be the residue of anatural or unnatural amino acid or a salt or ester thereof;

Q is hydrogen, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, orbenzoimidazolyl;

x is 0, 1, 2, 3, or 4;

G is a direct bond or oxygen, nitrogen, or sulfur;

z is 0, 1, 2, 3, 4, or 5;

m is 0 or 1;

R²⁴ is selected from a group consisting of hydrogen, alkyl,mercaptoalkyl, alkenyl, alkynyl, aroyl, alkylcarbonyl,aminoalkylcarbonyl, cycloalkyl, aryl, arylalkyl, thiazolyl, triazolyl,imidazolyl, benzothiazolyl, and benzoimidazolyl;

each R²³ is independently selected from hydrogen, halogen, cyano,hydroxyl, alkoxy, thiol, amino, nitro, alkyl, aryl, carbocyclic, orheterocyclic, and pharmaceutically acceptable salts, esters, andprodrugs thereof.

In one embodiment, R¹¹ is hydrogen. In another, A is oxygen. Forexample, n may be 3 and m may be 1. In other embodiments, R²⁴ ishydrogen or benzyl.

In certain embodiments, z is 0, 2, or 3. In others, R²⁵ is hydroxyl oralkoxy, e.g., methoxy, ethoxy, etc. In certain embodiments, two or moreR²⁵ substituents can be linked to form a fused ring (e.g., to form amethylendioxyphenyl moiety).

Examples of compounds of the invention include:

and pharmaceutically acceptable salts, esters, and prodrugs thereof.

In another embodiment, compounds of the invention include compound ofthe formula:

wherein:

R¹ is hydrogen, a substituted or unsubstituted cycloalkyl, heterocyclic,aryl, arylcycloalkyl, bicyclic or tricyclic ring, a bicyclic ortricyclic fused ring group, or a substituted or unsubstituted C₂-C₁₀alkyl group;

R² is selected from a group consisting of hydrogen, alkyl,mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl,triazolyl, imidazolyl, benzothiazolyl, and benzoimidazolyl;

Y is SO₃ ⁻X⁺, OSO₃ ⁻X⁺, or SSO₃ ⁻X⁺;

X⁺ is hydrogen, a cationic group, or an ester-forming group;

L¹ is a substituted or unsubstituted C₁-C₅ alkyl group or absent,

B is C₁-C₅ alkyl, alkenyl, or alkynyl group, optionally fused with Wwhen M is absent;

M is a covalent bond, amino, C₁-C₆ alkyl, alkenyl, alkynyl, carboxyl,oxy, amide, ester, thioether, thioester or absent;

W is a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl,aryl, arylalkyl, bicyclic or tricyclic ring, a bicyclic or tricyclicfused ring group, heterocyclic, thiazolyl, triazolyl, imidazolyl,benzothiazolyl, or benzoimidazolyl; and

v is 1, 2, 3, 4, 5, or 6; or a pharmaceutically acceptable salt, esteror prodrug thereof, provided that when Y is methyl, R¹ and R² arehydrogen, Y is SO₃ ⁻X⁺, M is a covalent bond, B is not CH₂—CH(M-W)—CH₂.

In a further embodiment, R¹ and R² are each hydrogen and L¹ is acovalent bond. In another embodiment, R¹ is alkyl and R² is hydrogen. Inanother further embodiment, Y is SO₃ ⁻X⁺. In another embodiment, v is 1.In another embodiment, M is a covalent bond or C₁-C₃ alkyl. In anotherembodiment, W is alkenyl. In another embodiment, W is aryl (e.g.,substituted or unsubstituted phenyl) or heteroaryl. In anotherembodiment, W is substituted or unsubstituted alkyl (e.g., straightchain, branched or cyclic (e.g., adamanyl, etc.).

In another embodiment, B is C₁-C₅ alkyl. Examples of B include:—CH(M-W)—CH₂—CH₂—, —CH₂—CH(M-W)—CH₂—, and —(CH₂)—CH₂—CH(M-W)—. Inanother embodiment B is alkenyl. In a further embodiment, the compoundis selected from the group consisting of:

In one embodiment, the invention pertains to compounds of Formula IX:

wherein:

R¹ is a substituted or unsubstituted cycloalkyl, heterocyclic, aryl,arylcycloalkyl, bicyclic or tricyclic ring, a bicyclic or tricyclicfused ring group, or a substituted or unsubstituted C₂-C₁₀ alkyl group;

R² is selected from the group consisting of hydrogen, alkyl,mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl, thiazolyl,triazolyl, imidazolyl, benzothiazolyl, and benzoimidazolyl;

R³ is hydrogen or a protecting group;

aa is a natural or unnatural amino acid residue;

L³ is a covalent bond, amino, C₁-C₆ alkyl, alkenyl, alkynyl, carboxyl,amide, aminoalkyl, ether, ester, thioether, thioester or absent;

Y is SO₃ ⁻X⁺, OSO₃ ⁻X⁺, or SSO₃ ⁻X⁺;

X⁺ is hydrogen, a cationic group, or ester-forming group; and

each of L¹ and L² is independently a substituted or unsubstituted C₁-C₅alkyl group or absent, or a pharmaceutically acceptable salt thereof.

In a further embodiment, R² is hydrogen, Y is SO₃ ⁻X⁺, and L² is—(CH₂)₃—. In another further embodiment, R¹ is carbocyclic orheterocyclic. In a further embodiment, R¹ is adamantyl. In a furtherembodiment, L³ is a covalent bond, a thioether, amino, oxy, aminoalkyl,or ether. In a further embodiment, R³ is hydrogen. In another furtherembodiment, aa is glycine, proline, alanine or phenylalanine. The R³moiety may be connected to the amino acid through any available atom,not necessarily through a peptide bond.

In a further embodiment, the compound of formula IX is selected from thegroup consisting of:

and pharmaceutically acceptable salts, esters and prodrugs thereof.

In another embodiment, the invention pertains to compounds of theformula (X):

wherein:

R^(a) is hydrogen, substituted or unsubstituted alkyl, aryl, heteroaryl,carboxyl, alkyloxycarbonyl, or aminocarbonyl;

R^(b) and R^(c) are each selected independently from hydrogen,substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, CONH₂,or R^(b), R^(c) and the carbon atom they are attached to can form asubstituted or unsubstituted cyclic structure of 4 to 8-membered ring ora fused ring system; and

X⁺ is hydrogen, a cationic group, or an ester-forming group, or apharmaceutically acceptable salt, ester, or prodrug thereof.

In a further embodiment, X⁺ is hydrogen.

In another further embodiment, R^(a) is substituted or unsubstitutedalkyl. Examples of R^(a) groups include methyl, ethyl, hydroxymethyl, orphenyl substituted alkyl (e.g., 1-(para-methyl-phenyl)-1-hydroxymethyl).In another embodiment, R^(a) is hydrogen or aminocarbonyl (e.g.,NH₂—C(═O)—).

In another embodiment, at least one of R^(b) and R^(c) are substitutedor unsubstituted alkyl. In another further embodiment, R^(b) and R^(c)are each unsubstituted alkyl. Examples of R^(b) and R^(c) includemethyl, ethyl, iso-propyl, propyl, iso-butyl, n-butyl, t-butyl, pentyl,hexyl, or heptyl. In another embodiment, at least one of R^(b) and R^(c)are hydroxyalkyl, alkoxyalkyl, alkylthioalkyl, aryloxyalkyl,alkylcarbonylalkyl, or arylalkyl.

In other embodiments, R^(b) and R^(c) are connected to form a ring. Thering may be cycloalkyl (e.g., cyclopropyl, cyclopentyl, cyclohexyl; orcycloheptyl) or cycloalkenyl. In other embodiments, R^(b) and R^(c) areconnected to form a bridged or fused ring system, e.g., adamantyl,norborane, indanyl, fluorenyl, etc.

Examples of compounds of Formula (X) include, but are not limited to:Compound N1, N3, N7, N8, N18, N28, N29, N44, N47, N49, N50, N51, N53,N56, N58, N59, N61, N62, N68, N72, N77, N81, MJ, NN, NM, NO, NP, NQ, NR,NT, NR, NZ, OC, OF, OG, ON, OQ, OS, OT, OU, OV, OW, OX, OY, PH, PJ, PK,PL, PM, PN, PO, PP, PR, PS, PT, PU, PV, PW, PX, PY, QB, QE, QF, and QM.

In another embodiment, the invention pertains to compounds of theformula (XI):

wherein:

R^(d) is H or alkyl;

R^(e) and R^(f) are each independently hydrogen, C₁-C₆ alkyl, or R^(e)and R^(f) taken together with the carbon they are attached to form a 3to 6-membered ring;

R^(g) is independently selected for each occurrence from the groupconsisting of: hydrogen, alkyl, alkoxy, halogen, NO₂, and alkyl-SO₂;

q is 1, 2, 3, 4, or 5;

X⁺ is hydrogen, a cationic group, or an ester-forming group;

Ar is aryl or heteroaryl; and

Z is 4-CH₂)₀₋₃—, —(CHOH)—, (CH₂)₁₋₃—O—(CH₂)₁₋₃, or a carbonyl group, ora pharmaceutically acceptable salt, ester, or prodrug thereof.

In one embodiment, X⁺ is hydrogen. In another embodiment, R^(d) ishydrogen. In yet another embodiment, R^(e) and R^(f) are eachindependently hydrogen, methyl, ethyl or are linked to form a ring,e.g., cyclohexyl ring. In another embodiment, Z is —CH₂—, —CHOH—, or acovalent bond. In another embodiment, Ar is phenyl, naphthyl,thiophenyl, furanyl, or benzothiophenyl. In yet another embodiment, q is1 or 2. Examples of R⁹ include methyl, ethyl, propyl, butyl, pentyl,hexyl, heptyl, hydroxy, bromine, chlorine, methoxy, ethoxy, propoxy,alkyl-SO₂—, and nitro.

Examples of compounds of Formula (XI) include, but are not limited to:Compound N37, N39, N43, N45, N46, N48, N52, N54, N55, N57, N60, N64,N65, N66, N67, NS, NU, NV, NX, NY, OA, OB, OD, OE, OH, OL, OM, OO, OP,OR, OZ, PA, PB, PD, PE, PF, PI, PQ, PZ, QA, QC, QD, QG, QH, QI, QJ, QK,QL and QW.

In another embodiment, the invention pertains to compounds of theformula (XII):

wherein:

R^(h) is hydrogen, benzyl, aryl-alkyl, aryl, or alkyl;

R^(i), R^(j), R^(k), R^(m), R^(n), and R^(o) are each independentlyhydrogen, substituted or unsubstituted aryl, substituted orunsubstituted benzyl, alkyl, alkenyl, carbocyclic, heterocyclic, absentor together may be linked to form a ring structure;

X⁺ is hydrogen, a cationic group, or an ester-forming group; and

t¹ and t² are each single or double bonds, provided that both t¹ and t²are not both double bonds, or a pharmaceutically acceptable salt, ester,or prodrug thereof.

In one embodiment, R^(h) is methyl, phenyl, indanyl, t-butyl, hydrogen,benzyl, or adamantyl. In a further embodiment, R^(i), R^(j), R^(k),R^(m), and R^(n) are each hydrogen and t¹ and t² are both single bonds.In another further embodiment, R^(o) is benzyl, phenyl, ethyl, butyl,thiophenyl-alkyl, or propylenyl.

In another embodiment, R^(j), R^(m), and R^(o) are each hydrogen, R^(n)and R^(k) are each absent, t¹ is a single bond, and t² is a double bond.

In yet another embodiment, R^(k), R^(m), R^(n) and R^(o) are eachhydrogen, and t¹ and t² are each single bonds.

In a further embodiment, R^(i) is benzyl, adamantyl, methyl, ethyl,propyl, butyl, pentyl, hexyl, heptyl, cyclohexyl, hydroxyalkyl, cubanyl,cubanyl-methyl-, adamantyl-methyl-, phenyl-adamantyl-methyl,aminocarbonyl-adamantyl-methyl-, or heteroaryl-alkyl.

In another further embodiment, R^(i), R^(j), R^(m), R^(n) and R^(o) areeach hydrogen, and t¹ and t² are each single bonds. In yet a furtherembodiment, R^(k) is benzyl, substituted phenyl, t-butyl, adamantyl,adamantyl-methyl, phenyl-adamantyl-methyl,aminocarbonyl-adamantyl-methyl-, or heteroaryl-alkyl.

In another embodiment, R^(i), R^(j), R^(k), and are each hydrogen andR^(m) and R^(o) are linked to form a ring. In a further embodiment, thering is unsubstituted or substituted cycloalkyl.

In another embodiment, the ring is a fused or a bridged ring.

Examples of compounds of Formula (XII) include, but are not limited to:Compound N2, N4, N5, N6, N9, N10, N11, N12, N13, N14, N15, N16, N17,N19, N20, N21, N22, N23, N24, N25, N26, N30, N32, N33, N34, N35, N36,N38, N40, N41, N42, N63, N69, N70, N71, N73, N74, N75, N76, N78, N79,N80, N82, N83, N84, N85, N86, N87, N88, QN, QO, QQ, QR, QS, QT, QU, QV,QX, QY, QZ, RA, RB, RC, RD, RE, RF, RG, RH, RI, RJ, RK, RL, RM, RN, RO,RP, RQ, RR, RS, RT, RU, RV, RW, RX, RY, RZ, SA, SB, SX, SY, SZ, TA andTB.

In another embodiment, the invention also pertains to compounds of theformula (XIII):

wherein:

n¹ is 0, 1, 2, or 3;

P is a covalent bond, alkyl, alkyloxy, amino, alkylamino, sulfur, oralkylthio;

X⁺ is hydrogen, a cationic group, or an ester-forming group; and

R^(p) is a natural or unnatural amino acid residue, or apharmaceutically acceptable salt, ester, or prodrug thereof.

In one embodiment, R^(p) is connected to P through a non-peptidic bond.The non-peptidic bond may originate from any carbon atom of the aminoacid residue. In certain embodiments, R^(p) may be connected to Pthrough a heteroatom. Examples of R^(p) include glycine (e.g.,HO(C═O)—CHNH—), phenylalanine, and proline. In another embodiment, P isa covalent bond, CH₂, —NH—, —O—, alkylthio, or alkyloxy.

Examples of compounds of Formula (XIII) include, but are not limited to:Compound SC, SD, SE, SF, SG, SH, SI, SJ, SK, SL, SM, SN, SO, SP, SQ, SR,SS, ST, SU, SV and SW.

In another embodiment, compound of the formula (XIV):

wherein:

n² is 0, 1, 2, or 3, selected such that three carbons are between theSO₃ ⁻X⁺ group and the nitrogen atom in the ring;

X⁺ is hydrogen, a cationic group, or an ester-forming group;

R^(s) is hydrogen or when n² is 3, R^(s) is (CH₂)₃—SO₃ ⁻X⁺;

R^(q) and R^(r) are each selected independently from hydrogen or alkyl,or a pharmaceutically acceptable salt, ester, or prodrug thereof.

In one embodiment, X⁺ is hydrogen.

In another embodiment, n² is 0 and the SO₃ ⁻X⁺ group is attached to atthe 4-position of the piperazine ring. In another embodiment, n² is 1and the SO₃ ⁻X⁺ group is attached to at the 3-position of the piperazinering. In yet another embodiment, n² is 2 and the SO₃ ⁻X⁺ group isattached to at the 2-position of the piperazine ring. In anotherembodiment, n² is 3 and R^(s) is (CH₂)₃—SO₃ ⁻X⁺.

Examples of compounds of Formula XIV include, but are not limited to:Compound OI, OJ, OK, PG and QP.

In yet another embodiment, the invention also pertains to compounds ofthe formula (XV):

wherein:

R^(t) is hydrogen, alkyl, or aryl;

R^(u) and R^(v) are each independently for each occurrence selected fromhydrogen, aryl, benzyl, alkyl, alkenyl, carbocyclic, heterocyclic, ortwo R^(u) or R^(v) groups on adjacent carbon atoms may form a doublebound, or together with the carbon atoms they are attached to forming acarbocyclic or heterocyclic ring;

n³ is 4, 5, 6, or 7; and

X⁺ is hydrogen, a cationic group, or an ester-forming group; or apharmaceutically acceptable salt, ester, or prodrug thereof.

Examples of compounds of Formula XV include, but are not limited to:Compound N27 and N31.

Other compounds of the invention include

and pharmaceutically acceptable salts, esters, and prodrugs thereof.

The invention pertains to both salt forms and acid/base forms of thecompounds of the invention. For example, the invention pertains not onlyto the particular salt forms of compounds shown herein as salts, butalso the invention includes other pharmaceutically acceptable salts, andthe acid and/or base form of the compound. The invention also pertainsto salt forms of compounds shown herein.

Compounds of the invention are also shown in Tables 2A, 2B, 3A, and 3Bbelow.

TABLE 2A ID STRUCTURE B

C

D

E

F

G

H

I

J

K

L

M

N

P

Q

R

S

X

Y

Z

AA

AB

AC

AD

AE

AF

AG

AH

AI

AJ

AK

AL

AM

AU

AV

AW

AX

AY

AZ

BA

BB

BC

BW

BX

BY

BZ

CC

CD

CE

CG

CH

CI

CJ

CK

CL

CM

CN

CO

CV

CY

DC

DD

DE

DG

DH

DI

DJ

DK

DL

DM

DN

DO

DP

DQ

DR

DS

DT

DU

DV

DW

DX

DY

DZ

EA

EB

EC

ED

EE

EF

EG

EH

EI

EJ

EK

EL

EN

EO

EP

EQ

ER

ES

ET

EV

EW

EY

EZ

FA

FH

FL

FM

FN

FO

FP

FQ

FR

FS

FT

FU

FV

FW

FX

FY

FZ

GA

GB

GC

GD

GE

GF

GH

GI

GJ

GK

GL

GM

GN

GO

GP

GQ

GR

GS

GT

GU

GZ

HA

HB

HC

HD

HE

HF

HG

HI

HJ

HK

HL

HM

HN

HO

HP

HQ

HR

HS

HT

HU

HV

HW

HX

HY

HZ

IA

IB

IC

ID

IE

IF

IG

IH

II

IJ

IK

IL

IM

IN

IO

IP

IR

IS

IT

IU

IV

IW

IX

IY

IZ

JA

JB

JC

JD

JE

JF

JG

JH

JI

JJ

JK

JL

JM

JN

JO

JP

JQ

JR

JS

JT

JU

JV

JW

JX

JY

JZ

KA

KB

KH

KI

KJ

KK

KL

KM

KN

KP

KQ

KR

KS

KT

KV

KW

KX

KY

LA

LC

LD

LE

LF

LG

LH

LI

LJ

LK

LL

LM

LN

LO

LP

LQ

NG

NH

NI

NJ

NK

NL

TABLE 2B P1

P2

P3

P4

P5

P6

P7

P8

P9

P10

P11

P12

P13

P14

P15

P16 NAO₃SOCH₂(CH₂)₃CH₂OSO₃Na P17

P178

P19

P20

P21

P22

P23

P24 HOCH₂CH₂CH₂CH₂SO₃Na P25 (NaO₃SCH₂CH₂CH₂CH₂)₂O P26

P27

P28

P29

P30

P31

P32

P33

P34

P35

P36 HC(CH₂OSO₃Na)₃ P37 CH₃C(CH₂OSO₃Na)₃ P38 NH₂CH₂CH₂CH₂SO₃Na P38

P40 NH₂C(CH₂OSO₃Na)₃ P41 NH₂CH₂CH₂OSO₃H P42

P43 NaO₃SNHCH₂CH₂OSO₃Na P44 H₂NCH₂CH₂CH₂OSO₃Na P45

P46 NaO₃SNHCH₂CH₂CH₂OSO₃Na P47 HN(CH₂CH₂OSO₃Na)₂ P48NaO₃SN(CH₂CH₂OSO₃Na)₂ P49

P50 H₂NCH₂CH₂SO₃H P51 H₂NCH₂CH₂SO₃H P52 NaO₃SOCH₂CH₂CH₂SO₃Na P53

P54

P55

P56

P57

P58

P59

P60

P61

P62

P63

P64 CH₃CH₂CH₂CH₂SO₃Na P65 CH₃(CH₂)₈CH₂SO₃Na P66

P67

P68 CH₃CH₂SO₃Na P69 CH₃CH₂CH₂SO₃Na P70 CH₃CH₂CH₂CH₂CH₂SO₃Na P71

P71

P73

P74

P75

P76

P77

P78

P79

P80

P81

P82

P823

P84

P85

P86

P87

P88

P89

P90

P91

P92

P93

P94

P95

P96

P97

P98

P99

P100

P101

P102

P103

P104

P105

P106

P107

P108

P109 CH₃(CH₂)₁₃N⁺(CH₃)₂[(CH₂)₃SO₃ ⁻] P110

P111

P112

P113

P114

P115

P116

P117

P118

P119

P120

P121

P122

P123 AcNHCH₂CH₂CH₂SO₃Na P124 BzNHCH₂CH₂CH₂SO₃Na P125

P126

P127 HOCH₂CH₂CH₂NHCH₂CH₂CH₂SO₃H P128

P129

P130

P131

P132

P134

P135

P136

P137

P138

P139

P140

P141

P142

P143

P144

P145

P146

P147

P148

P149

P150

P151

P152

P153

P154

P155

P156

P157

P158

P159

P160

P161

P162

P163

P164

P165

P166

P167

P168

P169

P170

P171

P172

P173

P174 CH₃(CH₂)₈NH(CH₂)₃SO₃H P175 CH₃(CH₂)₉NH(CH₂)₃SO₃H P176CH₃(CH₂)₁₀NH(CH₂)₃SO₃H P177 CH₃(CH₂)₁₁NH(CH₂)₃SO₃H P178CH₃(CH₂)₁₂NH(CH₂)₃SO₃H P179 CH₃(CH₂)₁₃NH(CH₂)₃SO₃H P180CH₃(CH₂)₁₅NH(CH₂)₃SO₃H P181 CH₃(CH₂)₁₇NHCH₂CH₂CH₂SO₃H P182

P183

P184

P185

P186

P187

P188

P189

P190

P191

P192

TABLE 3A ID STRUCTURE MJ

NN

MX

NE

NM

NO

NP

NQ

NR

NS

NT

NU

NV

NW

NX

NY

NZ

OA

OB

OC

OD

OE

OF

OG

OH

OI

OJ

OK

OL

OM

ON

OO

OP

OQ

OR

OS

OT

OU

OV

OW

OX

OY

OZ

PA

PB

PD

PE

PF

PG

PH

PI

PJ

PK

PL

PM

PN

PO

PP

PQ

PR

PS

PT

PU

PV

PW

PX

PY

PZ

QA

QB

QC

QD

QE

QF

QG

QH

QI

QJ

QK

QL

QM

QN

QO

QP

QQ

QR

QS

QT

QU

QV

QW

QX

QY

QZ

RA

RB

RC

RD

RE

RF

RG

RH

RI

RJ

RK

RL

RM

RN

RO

RP

RQ

RR

RS

RT

RU

RV

RW

RX

RY

RZ

SA

SB

SC

SD

SE

SF

SG

SH

SI

SJ

SK

SL

SM

SN

SO

SP

SQ

SR

SS

ST

SU

SV

SW

SX

SY

SZ

TA

TB

TABLE 3B ID Structure N1 

N2 

N3 

N4 

N5 

N6 

N7 

N8 

N9 

N10

N11

N12

N13

N14

N15

N16

N17

N18

N19

N20

N21

N22

N23

N24

N25

N26

N27

N28

N29

N30

N31

N32

N33

N34

N35

N36

N37

N38

N39

N40

N41

N42

N43

N44

N45

N46

N47

N48

N49

N50

N51

N52

N53

N54

N55

N56

N57

N58

N59

N60

N61

N62

N63

N64

N65

N66

N67

N68

N69

N70

N71

N72

N73

N74

N75

N76

N77

N78

N79

N80

N81

N82

N83

N84

N85

N86

N87

N88

N89

N90

N91

It should be noted that in the above table and throughout theapplication when an atom is shown without hydrogens, but hydrogens arerequired or chemically necessary to form a stable compound, hydrogensshould be inferred to be part of the compound.

In one embodiment, the invention does not pertain to the compoundsdescribed in WO 00/64420 and WO 96/28187. In this embodiment, theinvention does not pertain to methods of using the compounds describedin WO 00/64420 and WO 96/28187 for the treatment of diseases ordisorders described therein. In a further embodiment, the inventionpertains to methods of using the compounds described in WO 00/64420 andWO 96/28187 for methods described in this application, which are notdescribed in WO 00/64420 and WO 96/28187. Both of WO 00/64420 and WO96/28187 are incorporated by reference herein in their entirety. In afurther embodiment, this application does not pertain to the compoundsdescribed in U.S. application Ser. Nos. 10/871,512, 10/871,514, or10/871,365, all filed on Jun. 18, 2004, and incorporated herein byreference. In a further embodiment, the invention does not pertain tothe compounds of Tables 2A or 2B.

In another embodiment, the invention pertains to methods of theinvention which use and pharmaceutical compositions comprising thecompounds of Tables 2A, 2B, 3A or 3B. In another, the invention pertainsto methods of the invention which use and pharmaceutical compositionscomprising the compounds of Tables 2A, 2B, 3A, or 3B. In anotherembodiment, the compounds of the invention do not include the compoundsof Table 2A or 2B. In another embodiment, the compounds of the inventiondo not include the compounds of Table 2A or 2B.

It should be understood that the use of any of the compounds describedherein or in the applications identified in “The Related Applications”Section is within the scope of the present invention and is intended tobe encompassed by the present invention and each of the applications areexpressly incorporated herein at least for these purposes, and arefurthermore expressly incorporated for all other purposes.

Subjects and Patient Populations

The term “subject” includes living organisms in which amyloidosis canoccur, or which are susceptible to amyloid diseases, e.g., Alzheimer'sdisease, Down's syndrome, CAA, dialysis-related (β₂M) amyloidosis,secondary (AA) amyloidosis, primary (AL) amyloidosis, hereditaryamyloidosis, diabetes, etc. Examples of subjects include humans,chickens, ducks, peking ducks, geese, monkeys, deer, cows, rabbits,sheep, goats, dogs, cats, mice, rats, and transgenic species thereof.Administration of the compositions of the present invention to a subjectto be treated can be carried out using known procedures, at dosages andfor periods of time effective to modulate amyloid aggregation oramyloid-induced toxicity in the subject as further described herein. Aneffective amount of the therapeutic compound necessary to achieve atherapeutic effect may vary according to factors such as the amount ofamyloid already deposited at the clinical site in the subject, the age,sex, and weight of the subject, and the ability of the therapeuticcompound to modulate amyloid aggregation in the subject. Dosage regimenscan be adjusted to provide the optimum therapeutic response. Forexample, several divided doses may be administered daily or the dose maybe proportionally reduced as indicated by the exigencies of thetherapeutic situation.

In certain embodiments of the invention, the subject is in need oftreatment by the methods of the invention, and is selected for treatmentbased on this need. A subject in need of treatment is art-recognized,and includes subjects that have been identified as having a disease ordisorder related to amyloid-deposition or amyloidosis, has a symptom ofsuch a disease or disorder, or is at risk of such a disease or disorder,and would be expected, based on diagnosis, e.g., medical diagnosis, tobenefit from treatment (e.g., curing, healing, preventing, alleviating,relieving, altering, remedying, ameliorating, improving, or affectingthe disease or disorder, the symptom of the disease or disorder, or therisk of the disease or disorder).

In an exemplary aspect of the invention, the subject is a human. Forexample, the subject may be a human over 30 years old, human over 40years old, a human over 50 years old, a human over 60 years old, a humanover 70 years old, a human over 80 years old, a human over 85 years old,a human over 90 years old, or a human over 95 years old. The subject maybe a female human, including a postmenopausal female human, who may beon hormone (estrogen) replacement therapy. The subject may also be amale human. In another embodiment, the subject is under 40 years old.

A subject may be a human at risk for Alzheimer's disease, e.g., beingover the age of 40 or having a predisposition for Alzheimer's disease.Alzheimer's disease predisposing factors identified or proposed in thescientific literature include, among others, a genotype predisposing asubject to Alzheimer's disease; environmental factors predisposing asubject to Alzheimer's disease; past history of infection by viral andbacterial agents predisposing a subject to Alzheimer's disease; andvascular factors predisposing a subject to Alzheimer's disease. Asubject may also have one or more risk factors for cardiovasculardisease (e.g., atherosclerosis of the coronary arteries, anginapectoris, and myocardial infarction) or cerebrovascular disease (e.g.,atherosclerosis of the intracranial or extracranial arteries, stroke,syncope, and transient ischemic attacks), such as hypercholesterolemia,hypertension, diabetes, cigarette smoking, familial or previous historyof coronary artery disease, cerebrovascular disease, and cardiovasculardisease. Hypercholesterolemia typically is defined as a serum totalcholesterol concentration of greater than about 5.2 mmol/L (about 200mg/dL).

Several genotypes are believed to predispose a subject to Alzheimer'sdisease. These include the genotypes such as presenilin-1, presenilin-2,and amyloid precursor protein (APP) missense mutations associated withfamilial Alzheimer's disease, and α-2-macroglobulin and LRP-1 genotypes,which are thought to increase the risk of acquiring sporadic(late-onset) Alzheimer's disease. E. van Uden, et al., J. Neurosci.22(21), 9298-304 (2002); J. J. Goto, et al., J. Mol. Neurosci. 19(1-2),37-41 (2002). Another genetic risk factor for the development ofAlzheimer's disease are variants of ApoE, the gene that encodesapolipoprotein E (particularly the apoE4 genotype), a constituent of thelow-density lipoprotein particle. W J Strittmatter, et al., Annu. Rev.Neurosci. 19, 53-77 (1996). The molecular mechanisms by which thevarious ApoE alleles alter the likelihood of developing Alzheimer'sdisease are unknown, however the role of ApoE in cholesterol metabolismis consistent with the growing body of evidence linking cholesterolmetabolism to Alzheimer's disease. For example, chronic use ofcholesterol-lowering drugs such as statins has recently been associatedwith a lower incidence of Alzheimer's disease, and cholesterol-loweringdrugs have been shown to reduce pathology in APP transgenic mice. Theseand other studies suggest that cholesterol may affect APP processing.ApoE4 has been suggested to alter Aβ trafficking (in and out of thebrain), and favor retention of Aβ in the brain. ApoE4 has also beensuggested to favor APP processing toward Aβ formation. Environmentalfactors have been proposed as predisposing a subject to Alzheimer'sdisease, including exposure to aluminum, although the epidemiologicalevidence is ambiguous. In addition, prior infection by certain viral orbacterial agents may predispose a subject to Alzheimer's disease,including the herpes simplex virus and chlamydia pneumoniae. Finally,other predisposing factors for Alzheimer's disease can include riskfactors for cardiovascular or cerebrovascular disease, includingcigarette smoking, hypertension and diabetes. “At risk for Alzheimer'sdisease” also encompasses any other predisposing factors not listedabove or as yet identified and includes an increased risk forAlzheimer's disease caused by head injury, medications, diet, orlifestyle.

The methods of the present invention can be used for one or more of thefollowing: to prevent Alzheimer's disease, to treat Alzheimer's disease,or ameliorate symptoms of Alzheimer's disease, or to regulate productionof or levels of amyloid β (Aβ) peptides. In an embodiment, the humancarries one or more mutations in the genes that encode β-amyloidprecursor protein, presenilin-1 or presenilin-2. In another embodiment,the human carries the Apolipoprotein ε4 gene. In another embodiment, thehuman has a family history of Alzheimer's Disease or a dementia illness.In another embodiment, the human has trisomy 21 (Down's Syndrome). Inanother embodiment, the subject has a normal or low serum total bloodcholesterol level. In another embodiment, the serum total bloodcholesterol level is less than about 200 mg/dL, or less than about 180,and it can range from about 150 to about 200 mg/dL. In anotherembodiment, the total LDL cholesterol level is less than about 100mg/dL, or less than about 90 mg/dL and can range from about 30 to about100 mg/dL. Methods of measuring serum total blood cholesterol and totalLDL cholesterol are well known to those skilled in the art and forexample include those disclosed in WO 99/38498 at p. 11, incorporated byreference herein. Methods of determining levels of other sterols inserum are disclosed in H. Gylling, et al., “Serum Sterols During StanolEster Feeding in a Mildly Hypercholesterolemic Population”, J. LipidRes. 40: 593-600 (1999).

In another embodiment, the subject has an elevated serum total bloodcholesterol level. In another embodiment, the serum total cholesterollevel is at least about 200 mg/dL, or at least about 220 mg/dL and canrange from about 200 to about 1000 mg/dL. In another embodiment, thesubject has an elevated total LDL cholesterol level. In anotherembodiment, the total LDL cholesterol level is greater than about 100mg/dL, or even greater than about 110 mg/dL and can range from about 100to about 1000 mg/dL.

In another embodiment, the human is at least about 40 years of age. Inanother embodiment, the human is at least about 60 years of age. Inanother embodiment, the human is at least about 70 years of age. Inanother embodiment, the human is at least about 80 years of age. Inanother embodiment, the human is at least about 85 years of age. In oneembodiment, the human is between about 60 and about 100 years of age.

In still a further embodiment, the subject is shown to be at risk by adiagnostic brain imaging technique, for example, one that measures brainactivity, plaque deposition, or brain atrophy.

In still a further embodiment, the subject is shown to be at risk by acognitive test such as Clinical Dementia Rating (“CDR”), Alzheimer'sDisease Assessment Scale-Cognition (“ADAS-Cog”), Disability Assessmentfor Dementia (“DAD”) or Mini-Mental State Examination (“MMSE”). Thesubject may exhibit a below average score on a cognitive test, ascompared to a historical control of similar age and educationalbackground. The subject may also exhibit a reduction in score ascompared to previous scores of the subject on the same or similarcognition tests.

In determining the CDR, a subject is typically assessed and rated ineach of six cognitive and behavioural categories: memory, orientation,judgement and problem solving, community affairs, home and hobbies, andpersonal care. The assessment may include historical informationprovided by the subject, or preferably, a corroborator who knows thesubject well. The subject is assessed and rated in each of these areasand the overall rating, (0, 0.5, 1.0, 2.0 or 3.0) determined. A ratingof 0 is considered normal. A rating of 1.0 is considered to correspondto mild dementia. A subject with a CDR of 0.5 is characterized by mildconsistent forgetfulness, partial recollection of events and “benign”forgetfulness. In one embodiment the subject is assessed with a ratingon the CDR of above 0, of above about 0.5, of above about 1.0, of aboveabout 1.5, of above about 2.0, of above about 2.5, or at about 3.0.

Another test is the Mini-Mental State Examination (MMSE), as describedby Folstein “Mini-mental state. A practical method for grading thecognitive state of patients for the clinician.” J. Psychiatr. Res.12:189-198, 1975. The MMSE evaluates the presence of global intellectualdeterioration. See also Folstein “Differential diagnosis of dementia.The clinical process.” Psychiatr Clin North Am. 20:45-57, 1997. The MMSEis a means to evaluate the onset of dementia and the presence of globalintellectual deterioration, as seen in Alzheimer's disease andmulti-infart dementia. The MMSE is scored from 1 to 30. The MMSE doesnot evaluate basic cognitive potential, as, for example, the so-calledIQ test. Instead, it tests intellectual skills. A person of “normal”intellectual capabilities will score a “30” on the MMSE objective test(however, a person with a MMSE score of 30 could also score well below“normal” on an IQ test). See, e.g., Kaufer, J. Neuropsychiatry Clin.Neurosci. 10:55-63, 1998; Becke, Alzheimer Dis Assoc Disord. 12:54-57,1998; Ellis, Arch. Neurol. 55:360-365, 1998; Magni, Int. Psychogeriatr.8:127-134, 1996; Monsch, Acta Neurol. Scand. 92:145-150, 1995. In oneembodiment, the subject scores below 30 at least once on the MMSE. Inanother embodiment, the subject scores below about 28, below about 26,below about 24, below about 22, below about 20, below about 18, belowabout 16, below about 14, below about 12, below about 10, below about 8,below about 6, below about 4, below about 2, or below about 1.

Another means to evaluate cognition, particularly Alzheimer's disease,is the Alzheimer's Disease Assessment Scale (ADAS-Cog), or a variationtermed the Standardized Alzheimer's Disease Assessment Scale (SADAS). Itis commonly used as an efficacy measure in clinical drug trials ofAlzheimer's disease and related disorders characterized by cognitivedecline. SADAS and ADAS-Cog were not designed to diagnose Alzheimer'sdisease; they are useful in characterizing symptoms of dementia and area relatively sensitive indicator of dementia progression. (See, e.g.,Doraiswamy, Neurology 48:1511-1517, 1997; and Standish, J. Am. Geriatr.Soc. 44:712-716, 1996.) Annual deterioration in untreated Alzheimer'sdisease patients is approximately 8 points per year (See, eg., Raskind,M Prim. Care Companion J Clin Psychiatry 2000 August; 2(4):134-138).

The Disability Assessment for Dementia (“DAD”) scale has been developedto measure a patient's ability to perform the activities of daily living(Gélinas I et al. Development of a Functional Measure for Persons withAlzheimer's Disease: The Disability Assessment for Dementia. Am. J.Occupational Therapy. 1999; 53: 471-481). Activities of daily living maybe assessed according to self care (i.e., dressing and personal hygiene)and instrumental activities (e.g., housework, cooking, and usinghousehold devices). The objectives of the DAD scale includequantitatively measuring functional abilities in activities of dailyliving in individuals with cognitive impairments and to help delineateareas of cognitive deficits that may impair performance in activities ofdaily living. The DAD is administered through an interview with thecaregiver. It measures actual performance in activities of daily livingof the individual as observed over a 2 week period prior to theinterview. The scale assesses the following domains of activities:hygiene, dressing, telephoning, continence, eating, meal preparation,outing activities, finance and correspondence, medication use, leisureand housework. A total score is obtained by adding the rating for eachquestion and converting this total score out of 100. Higher scoresrepresent less disability in ADL while lower scores indicate moredysfunction. In one embodiment, the subject scores below 100 at leastonce on the DAD. In another embodiment, the subject scores below about95, below about 90, below about 85, below about 80, below about 75,below about 70, below about 65, below about 60, below about 55, belowabout 50, below about 45, below about 40, below about 30, below about20, or below about 10.

The ADAS-cog is designed to measure, with the use of questionnaires, theprogression and the severity of cognitive decline as seen in AD on a70-point scale. The ADAS-cog scale quantifies the number of wronganswers. Consequently, a high score on the scale indicates a more severecase of cognitive decline. In one embodiment, a subject exhibits a scoreof greater than 0, greater than about 5, greater than about 10, greaterthan about 15, greater than about 20, greater than about 25, greaterthan about 30, greater than about 35, greater than about 40, greaterthan about 45, greater than about 50, greater than about 55, greaterthan about 60, greater than about 65, greater than about 68, or about70.

In another embodiment, the subject exhibits no symptoms of Alzheimer'sDisease. In another embodiment, the subject is a human who is at least40 years of age and exhibits no symptoms of Alzheimer's Disease. Inanother embodiment, the subject is a human who is at least 40 years ofage and exhibits one or more symptoms of Alzheimer's Disease.

In another embodiment, the subject has Mild Cognitive Impairment. In afurther embodiment, the subject has a CDR rating of about 0.5. Inanother embodiment, the subject has early Alzheimer's disease. Inanother embodiment, the subject has cerebral amyloid angiopathy.

By using the methods of the present invention, the levels of amyloid βpeptides in a subject's plasma or cerebrospinal fluid (CSF) can bereduced from levels prior to treatment from about 10 to about 100percent, or even about 50 to about 100 percent.

In an alternative embodiment, the subject can have an elevated level ofamyloid Aβ₄₀ and Aβ₄₂ peptide in the blood and CSF prior to treatment,according to the present methods, of greater than about 10 pg/mL, orgreater than about 20 pg/mL, or greater than about 35 pg/mL, or evengreater than about 40 pg/mL. In another embodiment, the elevated levelof amyloid Aβ₄₂ peptide can range from about 30 pg/mL to about 200pg/mL, or even to about 500 pg/mL. One skilled in the art wouldunderstand that as Alzheimer's disease progresses, the measurable levelsof amyloid β peptide in the CSF may decrease from elevated levelspresent before onset of the disease. This effect is attributed toincreased deposition, i.e., trapping of Aβ peptide in the brain insteadof normal clearance from the brain into the CSF.

In an alternative embodiment, the subject can have an elevated level ofamyloid Aβ₄₀ peptide in the blood and CSF prior to treatment, accordingto the present methods, of greater than about 5 pg Aβ₄₂/mL or greaterthan about 50 pg Aβ₄₀/mL, or greater than about 400 pg/mL. In anotherembodiment, the elevated level of amyloid Aβ₄₀ peptide can range fromabout 200 pg/mL to about 800 pg/mL, to even about 1000 pg/mL.

In another embodiment, the subject can have an elevated level of amyloidAβ₄₂ peptide in the CSF prior to treatment, according to the presentmethods, of greater than about 5 pg/mL, or greater than about 10 pg/mL,or greater than about 200 pg/mL, or greater than about 500 pg/mL. Inanother embodiment, the level of amyloid β peptide can range from about10 pg/mL to about 1,000 pg/mL, or even about 100 pg/mL to about 1,000pg/mL.

In another embodiment, the subject can have an elevated level of amyloidAβ₄₀ peptide in the CSF prior to treatment according to the presentmethods of greater than about 10 pg/mL, or greater than about 50 pg/mL,or even greater than about 100 pg/mL. In another embodiment, the levelof amyloid β peptide can range from about 10 pg/mL to about 1,000 pg/mL.

The amount of amyloid a peptide in the brain, CSF, blood, or plasma of asubject can be evaluated by enzyme-linked immunosorbent assay (“ELISA”)or quantitative immunoblotting test methods or by quantitative SELDI-TOFwhich are well known to those skilled in the art, such as is disclosedby Zhang, et al., J. Biol. Chem. 274, 8966-72 (1999) and Zhang, et al.,Biochemistry 40, 5049-55 (2001). See also, A. K. Vehmas, et al., DNACell Biol. 20(11), 713-21 (2001), P. Lewczuk, et al., Rapid Commun. MassSpectrom. 17(12), 1291-96 (2003); B. M. Austen, et al., J. Peptide Sci.6, 459-69 (2000); and H. Davies, et al., BioTechniques 27, 1258-62(1999). These tests are performed on samples of the brain or blood whichhave been prepared in a manner well known to one skilled in the art.Another example of a useful method for measuring levels of amyloid apeptides is by Europium immunoassay (EIA). See, e.g., WO 99/38498 at p.11.

The methods of the invention may be applied as a therapy for a subjecthaving Alzheimer's disease or a dementia, or the methods of theinvention may be applied as a prophylaxis against Alzheimer's disease ordementia for subject with such a predisposition, as in a subject, e.g.,with a genomic mutation in the APP gene, the ApoE gene, or a presenilingene. The subject may have (or may be predisposed to developing or maybe suspected of having) vascular dementia, or senile dementia, MildCognitive Impairment, or early Alzheimer's disease. In addition toAlzheimer's disease, the subject may have another amyloid-relateddisease such as cerebral amyloid angiopathy, or the subject may haveamyloid deposits, especially amyloid-β amyloid deposits in the brain.

Treatment of Amyloid-Related Diseases

The present invention pertains to methods of using the compounds andpharmaceutical compositions thereof in the treatment and prevention ofamyloid-related diseases. The pharmaceutical compositions of theinvention may be administered therapeutically or prophylactically totreat diseases associated with amyloid (e.g., AL amyloid protein (λ orκ-chain related, e.g., amyloid λ, amyloid κ, amyloid κIV, amyloid λVI,amyloid γ, amyloid γ1), Aβ, IAPP, β₂M, AA, or AH amyloid protein) fibrilformation, aggregation or deposition.

The pharmaceutical compositions of the invention may act to amelioratethe course of an amyloid-related disease using any of the followingmechanisms (this list is meant to be illustrative and not limiting):slowing the rate of amyloid fibril formation or deposition; lesseningthe degree of amyloid deposition; inhibiting, reducing, or preventingamyloid fibril formation; inhibiting neurodegeneration or cellulartoxicity induced by amyloid; inhibiting amyloid induced inflammation;enhancing the clearance of amyloid from the brain; enhancing degradationof Aβ in the brain; or favoring clearance of amyloid protein prior toits organization in fibrils.

“Modulation” of amyloid deposition includes both inhibition, as definedabove, and enhancement of amyloid deposition or fibril formation. Theterm “modulating” is intended, therefore, to encompass prevention orstopping of amyloid formation or accumulation, inhibition or slowingdown of further amyloid formation or accumulation in a subject withongoing amyloidosis, e.g., already having amyloid deposition, andreducing or reversing of amyloid formation or accumulation in a subjectwith ongoing amyloidosis; and enhancing amyloid deposition, e.g.,increasing the rate or amount of amyloid deposition in vivo or in vitro.Amyloid-enhancing compounds may be useful in animal models ofamyloidosis, for example, to make possible the development of amyloiddeposits in animals in a shorter period of time or to increase amyloiddeposits over a selected period of time. Amyloid-enhancing compounds maybe useful in screening assays for compounds which inhibit amyloidosis invivo, for example, in animal models, cellular assays and in vitro assaysfor amyloidosis. Such compounds may be used, for example, to providefaster or more sensitive assays for compounds. Modulation of amyloiddeposition is determined relative to an untreated subject or relative tothe treated subject prior to treatment.

“Inhibition” of amyloid deposition includes preventing or stopping ofamyloid formation, e.g., fibrillogenesis, clearance of amyloid, e.g.,soluble Aβ from brain, inhibiting or slowing down of further amyloiddeposition in a subject with amyloidosis, e.g., already having amyloiddeposits, and reducing or reversing amyloid fibrillogenesis or depositsin a subject with ongoing amyloidosis. Inhibition of amyloid depositionis determined relative to an untreated subject, or relative to thetreated subject prior to treatment, or, e.g., determined by clinicallymeasurable improvement, e.g., or in the case of a subject with brainamyloidosis, e.g., an Alzheimer's or cerebral amyloid angiopathysubject, stabilization of cognitive function or prevention of a furtherdecrease in cognitive function (i.e., preventing, slowing, or stoppingdisease progression), or improvement of parameters such as theconcentration of Aβ or tau in the CSF.

As used herein, “treatment” of a subject includes the application oradministration of a composition of the invention to a subject, orapplication or administration of a composition of the invention to acell or tissue from a subject, who has an amyloid-related disease orcondition, has a symptom of such a disease or condition, or is at riskof (or susceptible to) such a disease or condition, with the purpose ofcuring, healing, alleviating, relieving, altering, remedying,ameliorating, improving, or affecting the disease or condition, thesymptom of the disease or condition, or the risk of (or susceptibilityto) the disease or condition. The term “treating” refers to any indiciaof success in the treatment or amelioration of an injury, pathology orcondition, including any objective or subjective parameter such asabatement; remission; diminishing of symptoms or making the injury,pathology or condition more tolerable to the subject; slowing in therate of degeneration or decline; making the final point of degenerationless debilitating; improving a subject's physical or mental well-being;or, in some situations, preventing the onset of dementia. The treatmentor amelioration of symptoms can be based on objective or subjectiveparameters; including the results of a physical examination, apsychiatric evaluation, or a cognition test such as CDR, MMSE, DAD,ADAS-Cog, or another test known in the art. For example, the methods ofthe invention successfully treat a subject's dementia by slowing therate of or lessening the extent of cognitive decline.

In one embodiment, the term “treating” includes maintaining a subject'sCDR rating at its base line rating or at 0. In another embodiment, theterm treating includes decreasing a subject's CDR rating by about 0.25or more, about 0.5 or more, about 1.0 or more, about 1.5 or more, about2.0 or more, about 2.5 or more, or about 3.0 or more. In anotherembodiment, the term “treating” also includes reducing the rate of theincrease of a subject's CDR rating as compared to historical controls.In another embodiment, the term includes reducing the rate of increaseof a subject's CDR rating by about 5% or more, about 10% or more, about20% or more, about 25% or more, about 30% or more, about 40% or more,about 50% or more, about 60% or more, about 70% or more, about 80% ormore, about 90% or more, or about 100%, of the increase of thehistorical or untreated controls.

In another embodiment, the term “treating” also includes maintaining asubject's score on the MMSE. The term “treating” includes increasing asubject's MMSE score by about 1, about 2, about 3, about 4, about 5,about 7.5, about 10, about 12.5, about 15, about 17.5, about 20, orabout 25 points. The term also includes reducing the rate of thedecrease of a subject's MMSE score as compared to historical controls.In another embodiment, the term includes reducing the rate of decreaseof a subject's MMSE score by about 5% or less, about 10% or less, about20% or less, about 25% or less, about 30% or less, about 40% or less,about 50% or less, about 60% or less, about 70% or less, about 80% orless, about 90% or less or about 100% or less, of the decrease of thehistorical or untreated controls.

In another embodiment, the term “treating” also includes maintaining asubject's score on the DAD. The term “treating” includes increasing asubject's DAD score by about 1, about 5, about 10, about 15, about 20,about 30, about 35, about 40, about 50, about 60, about 70, or about 80points. The term also includes reducing the rate of the decrease of asubject's DAD score as compared to historical controls. In anotherembodiment, the term includes reducing the rate of decrease of asubject's DAD score by about 5% or less, about 10% or less, about 20% orless, about 25% or less, about 30% or less, about 40% or less, about 50%or less, about 60% or less, about 70% or less, about 80% or less, about90% or less or about 100% or less, of the decrease of the historical oruntreated controls.

In yet another embodiment, the term “treating” includes maintaining asubject's score on the ADAS-Cog. The term “treating” includes decreasinga subject's ADAS-Cog score by about 1 point or greater, by about 2points or greater, by about 3 points or greater, by about 4 points orgreater, by about 5 points or greater, by about 7.5 points or greater,by about 10 points or greater, by about 12.5 points or greater, by about15 points or greater, by about 17.5 points or greater, by about 20points or greater, or by about 25 points or greater. The term alsoincludes reducing the rate of the increase of a subject's ADAS-Cog scoreas compared to historical controls. In another embodiment, the termincludes reducing the rate of increase of a subject's ADAS-Cog score byabout 5% or more, about 10% or more, about 20% or more, about 25% ormore, about 30% or more, about 40% or more, about 50% or more, about 60%or more, about 70% or more, about 80% or more, about 90% or more orabout 100% of the increase of the historical or untreated controls.

In another embodiment, the term “treating” e.g., for AA or ALamyloidosis, includes an increase in serum creatinine, e.g., an increaseof creatinine clearance of 10% or greater, 20% or greater, 50% orgreater, 80% or greater, 90% or greater, 100% or greater, 150% orgreater, 200% or greater. The term “treating” also may induce remissionof nephrotic syndrome (NS). It may also include remission of chronicdiarrhea and/or a gain in body weight, e.g., by 10% or greater, 15% orgreater, or 20% or greater.

Without wishing to be bound by theory, in some aspects thepharmaceutical compositions of the invention contain a compound thatprevents or inhibits amyloid fibril formation, either in the brain orother organ of interest (acting locally) or throughout the entire body(acting systemically). Pharmaceutical compositions of the invention maybe effective in controlling amyloid deposition either following theirentry into the brain (following penetration of the blood brain barrier)or from the periphery. When acting from the periphery, a compound of apharmaceutical composition may alter the equilibrium of amyloidogenicpeptide between the brain and the plasma so as to favor the exit ofamyloidogenic peptide from the brain. It may also favor clearance (orcatabolism) of the amyloid protein (soluble), and then prevent amyloidfibril formation and deposition due to a reduction of the amyloidprotein pool in a specific organ, e.g., liver, spleen, pancreas, kidney,joints, brain, etc. An increase in the exit of amyloidogenic peptidefrom the brain would result in a decrease in amyloidogenic peptide brainconcentration and therefore favor a decrease in amyloidogenic peptidedeposition. In particular, an agent may lower the levels of amyloid βpeptides, e.g., both Aβ40 and Aβ42 in the CSF and the plasma, or theagent may lower the levels of amyloid β peptides, e.g., Aβ40 and Aβ42 inthe CSF and increase it in the plasma. Alternatively, compounds thatpenetrate the brain could control deposition by acting directly on brainamyloidogenic peptide e.g., by maintaining it in a non-fibrillar form orfavoring its clearance from the brain, by increasing its degradation inthe brain, or protecting brain cells from the detrimental effect ofamyloidogenic peptide. An agent can also cause a decrease of theconcentration of the amyloid protein (i.e., in a specific organ so thatthe critical concentration needed to trigger amyloid fibril formation ordeposition is not reached). Furthermore, the compounds described hereinmay inhibit or reduce an interaction between amyloid and a cell surfaceconstituent, for example, a glycosaminoglycan or proteoglycanconstituent of a basement membrane, whereby inhibiting or reducing thisinteraction produces the observed neuroprotective and cell-protectiveeffects. For example, the compound may also prevent an amyloid peptidefrom binding or adhering to a cell surface, a process which is known tocause cell damage or toxicity. Similarly, the compound may blockamyloid-induced cellular toxicity or microglial activation oramyloid-induced neurotoxicity, or inhibit amyloid induced inflammation.The compound may also reduce the rate or amount of amyloid aggregation,fibril formation, or deposition, or the compound lessens the degree ofamyloid deposition. The foregoing mechanisms of action should not beconstrued as limiting the scope of the invention inasmuch as theinvention may be practiced without such information.

The Aβ peptide has been shown by several groups to be highly toxic toneurons. Amyloid plaques are directly associated with reactive gliosis,dystrophic neurites and apoptotic cells, suggesting that plaques induceneurodegenerative changes. Neurotoxicity may eventually disrupt or evenkill neurons. In vitro, Aβ has been shown to induce apoptosis in manydifferent neuronal cell types, such as rat PC-12 cells, primary rathippocampal and cortical cultures, and the predifferentiated humanneurotype SH-SY5Y cell line (Dickson D W (2004) J Clin Invest 114:23-7;Canu et al. (2003) Cerebellum 2:270-278; Li et al. (1996) Brain Research738:196-204). Numerous reports have shown that Aβ fibrils can induceneurodegeneration, and it has been shown that neuronal cells exposed invitro to Aβ can become apoptotic (Morgan et al. (2004) Prog. Neurobiol.74:323-349; Stefani et al. (2003) J. Mol. Med. 81:678-99; La Ferla etal. (1997) J. Clin. Invest. 100(2):310-320). In Alzheimer's disease, aprogressive neuronal cell loss accompanies the deposition of Aβ amyloidfibrils in senile plaques.

In yet another aspect, the invention pertains to a method for inhibitingAβ-induced neuronal cell death by administering an effective amount of acompound of the present invention.

Another aspect of the invention pertains to a method for providingneuroprotection to a subject having an Aβ-amyloid related disease, e.g.Alzheimer's disease, that includes administering an effective amount ofa compound of the present invention to the subject, such thatneuroprotection is provided.

In another aspect, methods for inhibiting Aβ-induced neuronal cell deathare provided that include administration of an effective amount of acompound of the present invention to a subject such that neuronal celldeath is inhibited.

In another aspect, methods for treating a disease state characterized byAβ-induced neuronal cell death in a subject are provided, e.g., byadministering an effective amount of a compound of the presentinvention. Non-limiting examples of such disease states includeAlzheimer's disease and Aβ-amyloid related diseases.

The term “neuroprotection” includes protection of neuronal cells of asubject from Aβ-induced cell death, e.g., cell death induced directly orindirectly by an Aβ peptide. Aβ-induced cell death may result ininitiation of processes such as, for example: the destabilization of thecytoskeleton; DNA fragmentation; the activation of hydrolytic enzymes,such as phospholipase A2; activation of caspases, calcium-activatedproteases and/or calcium-activated endonucleases; inflammation mediatedby macrophages; calcium influx into a cell; membrane potential changesin a cell; the disruption of cell junctions leading to decreased orabsent cell-cell communication; and the activation of expression ofgenes involved in cell death, e.g., immediate-early genes.

The term “amyloid-β disease” (or “amyloid-β related disease,” whichterms as used herein are synonymous) may be used for mild cognitiveimpairment; vascular dementia; early Alzheimer's disease; Alzheimer'sdisease, including sporadic (non-hereditary) Alzheimer's disease andfamilial (hereditary) Alzheimer's disease; age-related cognitivedecline; cerebral amyloid angiopathy (“CAA”); hereditary cerebralhemorrhage; senile dementia; Down's syndrome; inclusion body myositis(“IBM”); or age-related macular degeneration (“ARMD”). According tocertain aspects of the invention, amyloid-β is a peptide having 39-43amino-acids, or amyloid-β is an amyloidogenic peptide produced fromPAPP.

Mild cognitive impairment (“MCI”) is a condition characterized by astate of mild but measurable impairment in thinking skills, which is notnecessarily associated with the presence of dementia. MCI frequently,but not necessarily, precedes Alzheimer's disease. It is a diagnosisthat has most often been associated with mild memory problems, but itcan also be characterized by mild impairments in other thinking skills,such as language or planning skills. However, in general, an individualwith MCI will have more significant memory lapses than would be expectedfor someone of their age or educational background. As the conditionprogresses, a physician may change the diagnosis to “Mild-to-ModerateCognitive Impairment,” as is well understood in this art.

Cerebral amyloid angiopathy (“CAA”) refers to the specific deposition ofamyloid fibrils in the walls of leptomingeal and cortical arteries,arterioles and in capillaries and veins. It is commonly associated withAlzheimer's disease, Down's syndrome and normal aging, as well as with avariety of familial conditions related to stroke or dementia (seeFrangione, et al., Amyloid: J. Protein Folding Disord. 8, Suppl. 1,36-42 (2001)). CAA can occur sporadically or be hereditary. Multiplemutation sites in either Aβ or the APP gene have been identified and areclinically associated with either dementia or cerebral hemorrhage.Exemplary CAA disorders include, but are not limited to, hereditarycerebral hemorrhage with amyloidosis of Icelandic type (HCHWA-I); theDutch variant of HCHWA (HCHWA-D; a mutation in Aβ); the Flemish mutationof Aβ; the Arctic mutation of Aβ; the Italian mutation of Aβ; the Iowamutation of Aβ; familial British dementia; and familial Danish dementia.Cerebral amyloid angiopathy is known to be associated with cerebralhemorrhage (or hemorrhagic stroke).

Additionally, abnormal accumulation of APP and of amyloid-β protein inmuscle fibers has been implicated in the pathology of sporadic inclusionbody myositis (“IBM”) (Askanas, et al., Proc. Natl. Acad. Sci. USA 93,1314-19 (1996); Askanas, et al., Current Opinion in Rheumatology 7,486-96 (1995)). Accordingly, the compounds of the invention can be usedprophylactically or therapeutically in the treatment of disorders inwhich amyloid-β protein is abnormally deposited at non-neurologicallocations, such as treatment of IBM by delivery of the compounds tomuscle fibers.

Additionally, it has been shown that Aβ is associated with abnormalextracellular deposits, known as drusen, that accumulate along the basalsurface of the retinal pigmented epithelium in individuals withage-related macular degeneration (ARMD). ARMD is a cause of irreversiblevision loss in older individuals. It is believed that Aβ depositioncould be an important component of the local inflammatory events thatcontribute to atrophy of the retinal pigmented epithelium, drusenbiogenesis, and the pathogenesis of ARMD (Johnson, et al., Proc. Natl.Acad. Sci. USA 99(18), 11830-5 (2002)). Therefore, the invention alsorelates to the treatment or prevention of age-related maculardegeneration.

Also, the invention relates to a method for preventing or inhibitingamyloid deposition in a subject. For example, such a method comprisesadministering to a subject a therapeutically effective amount of acompound capable of reducing the concentration of amyloid (e.g., ALamyloid protein (λ or κ-chain related, e.g., amyloid λ, amyloid κ,amyloid κIV, amyloid λVI, amyloid γ, amyloid γ1) Aβ, IAPP, β₂M, AA, AHamyloid protein, or other amyloids), such that amyloid accumulation ordeposition is prevented or inhibited.

In another aspect, the invention relates to a method for preventing,reducing, or inhibiting amyloid deposition in a subject. For example,such a method comprises administering to a subject a therapeuticallyeffective amount of a compound capable of inhibiting amyloid (e.g., ALamyloid protein (λ or κ-chain related, e.g., amyloid λ, amyloid κ,amyloid κIV, amyloid λVI, amyloid γ, amyloid γ1) Aβ, IAPP, β₂M, AA, AHamyloid protein, or other amyloids), such that amyloid deposition isprevented, reduced, or inhibited.

The invention also relates to a method for modulating, e.g., minimizing,amyloid-associated damage to cells, comprising the step of administeringa compound capable of reducing the concentration of amyloid (e.g., ALamyloid protein (λ or κ-chain related, e.g., amyloid λ, amyloid κ,amyloid κIV, amyloid λVI, amyloid γ, amyloid γ1) Aβ, IAPP, β₂M, AA, AHamyloid protein, or another amyloid), such that said amyloid-associateddamage to cells is modulated. In certain aspects of the invention, themethods for modulating amyloid-associated damage to cells comprise astep of administering a compound capable of reducing the concentrationof amyloid or reducing the interaction of an amyloid with a cellsurface.

The invention also includes a method for directly or indirectlypreventing cell death in a subject, the method comprising administeringto a subject a therapeutically effective amount of a compound capable ofpreventing amyloid (e.g., AL amyloid protein (λ or κ-chain related,e.g., amyloid λ, amyloid κ, amyloid κIV, amyloid λVI, amyloid γ, amyloidγ1), Aβ, IAPP, β₂M, AA, AH amyloid protein, or other amyloid) mediatedevents that lead, directly or indirectly, to cell death.

In an embodiment, the method is used to treat Alzheimer's disease (e.g.sporadic or familial AD). The method can also be used prophylacticallyor therapeutically to treat other clinical occurrences of amyloid-(3deposition, such as in Down's syndrome individuals and in patients withcerebral amyloid angiopathy (“CAA”) or hereditary cerebral hemorrhage.

The compounds of the invention may be used prophylactically ortherapeutically in the treatment of disorders in which amyloid-betapeptide is abnormally deposited at non-neurological locations, such astreatment of IBM by delivery of the compounds to muscle fibers, ortreatment of macular degeneration by delivery of the compound(s) of theinvention to the basal surface of the retinal pigmented epithelium.

The present invention also provides a method for modulatingamyloid-associated damage to cells, comprising the step of administeringa compound capable of reducing the concentration of Aβ, or capable ofminimizing the interaction of Aβ (soluble oligomeric or fibrillary) withthe cell surface, such that said amyloid-associated damage to cells ismodulated. In certain aspects of the invention, the methods formodulating amyloid-associated damage to cells comprise a step ofadministering a compound capable of reducing the concentration of Aβ orreducing the interaction of Aβ with a cell surface.

In accordance with the present invention, there is further provided amethod for preventing cell death in a subject, said method comprisingadministering to a subject a therapeutically effective amount of acompound capable of preventing Aβ-mediated events that lead, directly orindirectly, to cell death.

The present invention also provides a method for modulatingamyloid-associated damage to cells, comprising the step of administeringa compound capable of reducing the concentration of IAPP, or capable ofminimizing the interaction of IAPP (soluble oligomeric or fibrillary)with the cell surface, such that said amyloid-associated damage to cellsis modulated. In certain aspects of the invention, the methods formodulating amyloid-associated damage to cells comprise a step ofadministering a compound capable of reducing the concentration of IAPPor reducing the interaction of IAPP with a cell surface.

In accordance with the present invention, there is further provided amethod for preventing cell death in a subject, said method comprisingadministering to a subject a therapeutically effective amount of acompound capable of preventing IAPP-mediated events that lead, directlyor indirectly, to cell death.

This invention also provides methods and compositions which are usefulin the treatment of amyloidosis. The methods of the invention involveadministering to a subject a therapeutic compound which inhibits amyloiddeposition. Accordingly, the compositions and methods of the inventionare useful for inhibiting amyloidosis in disorders in which amyloiddeposition occurs. The methods of the invention can be usedtherapeutically to treat amyloidosis or can be used prophylactically ina subject susceptible to (hereditary) amyloidosis or identified as beingat risk to develop amyloidosis, e.g., hereditary, or identified as beingat risk to develop amyloidosis. In certain embodiments, the inventionincludes a method of inhibiting an interaction between an amyloidogenicprotein and a constituent of basement membrane to inhibit amyloiddeposition. The constituent of basement membrane is a glycoprotein orproteoglycan, preferably heparan sulfate proteoglycan. A therapeuticcompound used in this method may interfere with binding of a basementmembrane constituent to a target binding site on an amyloidogenicprotein, thereby inhibiting amyloid deposition.

In some aspects, the methods of the invention involve administering to asubject a therapeutic compound which inhibits amyloid deposition.“Inhibition of amyloid deposition,” includes the prevention of amyloidformation, inhibition of further amyloid deposition in a subject withongoing amyloidosis and reduction of amyloid deposits in a subject withongoing amyloidosis. Inhibition of amyloid deposition is determinedrelative to an untreated subject or relative to the treated subjectprior to treatment. In an embodiment, amyloid deposition is inhibited byinhibiting an interaction between an amyloidogenic protein and aconstituent of basement membrane. “Basement membrane” refers to anextracellular matrix comprising glycoproteins and proteoglycans,including laminin, collagen type IV, fibronectin, perlecan, agrin,dermatan sulfate, and heparan sulfate proteoglycan (HSPG). In oneembodiment, amyloid deposition is inhibited by interfering with aninteraction between an amyloidogenic protein and a sulfatedglycosaminoglycan such as HSPG, dermatan sulfate, perlecan or agrinsulfate. Sulfated glycosaminoglycans are known to be present in alltypes of amyloids (see Snow, et al. Lab. Invest. 56, 120-23 (1987)) andamyloid deposition and HSPG deposition occur coincidentally in animalmodels of amyloidosis (see Snow, et al. Lab. Invest. 56, 665-75 (1987)and Gervais, F. et al. Curr. Med. Chem., 3, 361-370 (2003)). Consensusbinding site motifs for HSPG in amyloidogenic proteins have beendescribed (see, e.g., Cardin and Weintraub Arteriosclerosis 9, 21-32(1989)). The ability of a compound to prevent or block the formation ordeposition of amyloid may reside in its ability to bind tonon-fibrillar, soluble amyloid protein and to maintain its solubililty.

The ability of a therapeutic compound of the invention to inhibit aninteraction between an amyloidogenic protein and a glycoprotein orproteoglycan constituent of a basement membrane can be assessed by an invitro binding assay, such as that described in U.S. Pat. No. 5,164,295,the contents of which are hereby incorporated by reference.Alternatively, the ability of a compound to bind to an amyloidogenicprotein or to inhibit the binding of a basement membrane constituent(e.g. HSPG) to an amyloidogenic protein (e.g. Aβ) can be measured usinga mass spectrometry assay where soluble protein, e.g. Aβ, IAPP, β₂M isincubated with the compound. A compound which binds to, e.g. Aβ, willinduce a change in the mass spectrum of the protein. Exemplary protocolsfor a mass spectrometry assay employing Aβ and IAPP can be found in theExamples, the results of which are provided in Table 3. The protocol canreadily be modified to adjust the sensitivity of the data, e.g., byadjusting the amount of protein and/or compound employed. Thus, e.g.,binding might be detected for test compounds noted as not havingdetectable binding employing less sensitive test protocols.

Alternative methods for screening compounds exist and can readily beemployed by a skilled practitioner to provide an indication of theability of test compounds to bind to, e.g., fibrillar Aβ. One suchscreening assay is an ultraviolet absorption assay. In an exemplaryprotocol, a test compound (20 μM) is incubated with 50 μM Aβ (1-40)fibers for 1 hour at 37° C. in Tris buffered saline (20 mM Tris, 150 mMNaCl, pH 7.4 containing 0.01 sodium azide). Following incubation, thesolution is centrifuged for 20 minutes at 21,000 g to sediment the Aβ(1-40) fibers along with any bound test compound. The amount of testcompound remaining in the supernatant can then be determined by readingthe absorbance. The fraction of test compound bound can then becalculated by comparing the amount remaining in the supernatants ofincubations with Aβ to the amount remaining in control incubations whichdo not contain Aβ fibers. Thioflavin T and Congo Red, both of which areknown to bind to Aβ fibers, may be included in each assay run aspositive controls. Before assaying, test compounds can be diluted to 40μM, which would be twice the concentration in the final test, and thenscanned using the Hewlett Packard 8453 UV/VIS spectrophotometer todetermine if the absorbance is sufficient for detection.

In another embodiment, the invention pertains to a method for improvingcognition in a subject suffering from an amyloid-related disease. Themethod includes administering an effective amount of a therapeuticcompound of the invention, such that the subject's cognition isimproved. The subject's cognition can be tested using methods known inthe art such as the Clinical Dementia Rating (“CDR”), Mini-Mental StateExamination (“MMSE”), Disability Assessment for Dementia (“DAD”), andthe Alzheimer's Disease Assessment Scale-Cognition (“ADAS-Cog”).

In another embodiment, the invention pertains to a method for treating asubject for an amyloid-related disease. The method includesadministering a cognitive test to a subject prior to administration of acompound of the invention, administering an effective amount of acompound of the invention to the subject, and administering a cognitivetest to the subject subsequent to administration of the compound, suchthat the subject is treated for the amyloid-related disease, wherein thesubject's score on said cognitive test is improved.

“Improvement,” “improved” or “improving” in cognition is present withinthe context of the present invention if there is a statisticallysignificant difference in the direction of normality between theperformance of subjects treated using the methods of the invention ascompared to members of a placebo group, historical control, or betweensubsequent tests given to the same subject.

In one embodiment, a subject's CDR is maintained at 0. In anotherembodiment, a subject's CDR is decreased (e.g., improved) by about 0.25or more, about 0.5 or more, about 1.0 or more, about 1.5 or more, about2.0 or more, about 2.5 or more, or about 3.0 or more. In anotherembodiment, the rate of increase of a subject's CDR rating is reduced byabout 5% or more, about 10% or more, about 20% or more, about 25% ormore, about 30% or more, about 40% or more, about 50% or more, about 60%or more, about 70% or more, about 80% or more, about 90% or more, orabout 100% or more of the increase of the historical or untreatedcontrols.

In one embodiment, a subject's score on the MMSE is maintained.Alternatively, the subject's score on the MMSE may be increased by about1, about 2, about 3, about 4, about 5, about 7.5, about 10, about 12.5,about 15, about 17.5, about 20, or about 25 points. In anotheralternative, the rate of the decrease of a subject's MMSE score ascompared to historical controls is reduced. For example, the rate of thedecrease of a subject's MMSE score may be reduced by about 5% or more,about 10% or more, about 20% or more, about 25% or more, about 30% ormore, about 40% or more, about 50% or more, about 60% or more, about 70%or more, about 80% or more, about 90% or more, or about 100% or more ofthe decrease of the historical or untreated controls.

In one embodiment, a subject's score on the DAD is maintained.Alternatively, the subject's score on the DAD may be increased by about1, about 2, about 3, about 4, about 5, about 7.5, about 10, about 15,about 20, about 30, about 40, or about 50 or more points. In anotheralternative, the rate of the decrease of a subject's DAD score ascompared to historical controls is reduced. For example, the rate of thedecrease of a subject's DAD score may be reduced by about 5% or more,about 10% or more, about 20% or more, about 25% or more, about 30% ormore, about 40% or more, about 50% or more, about 60% or more, about 70%or more, about 80% or more, about 90% or more, or about 100% or more ofthe decrease of the historical or untreated controls.

In one embodiment, the invention pertains to a method for treating,slowing or stopping an amyloid-related disease associated with cognitiveimpairment, by administering to a subject an effective amount of atherapeutic compound of the invention, wherein the annual deteriorationof the subject's cognition as measured by ADAS-Cog is less than 8 pointsper year, less the 6 points per year, less than 5 points per year, lessthan 4 points per year, or less than 3 points per year. In a furtherembodiment, the invention pertains to a method for treating, slowing orstopping an amyloid-related disease associated with cognition byadministering an effective amount of a therapeutic compound of theinvention such that the subject's cognition as measured by ADAS-Cogremains constant over a year. “Constant” includes fluctuations of nomore than 2 points. Remaining constant includes fluctuations of twopoints or less in either direction. In a further embodiment, thesubject's cognition improves by 2 points or greater per year, 3 pointsor greater per year, 4 point or greater per year, 5 points or greaterper year, 6 points or greater per year, 7 points or greater per year, 8points or greater per year, etc. as measured by the ADAS-Cog. In anotheralternative, the rate of the increase of a subject's ADAS-Cog score ascompared to historical controls is reduced. For example, the rate of theincrease of a subject's ADAS-Cog score may be reduced by about 5% ormore, about 10% or more, about 20% or more, about 25% or more, about 30%or more, about 40% or more, about 50% or more, about 60% or more, about70% or more, about 80% or more, about 90% or more or about 100% of theincrease of the historical or untreated controls.

In another embodiment, the ratio of Aβ42:Aβ40 in the CSF or plasma of asubject decreases by about 15% or more, about 20% or more, about 25% ormore, about 30% or more, about 35% or more, about 40% or more, about 45%or more, or about 50% or more. In another embodiment, the levels of Aβin the subject's cerebrospinal fluid decrease by about 15% or more,about 25% or more, about 35% or more, about 45% or more, about 55% ormore, about 75% or more, or about 90% or more.

It is to be understood that wherever values and ranges are providedherein, e.g., in ages of subject populations, dosages, and blood levels,all values and ranges encompassed by these values and ranges, are meantto be encompassed within the scope of the present invention. Moreover,all values in these values and ranges may also be the upper or lowerlimits of a range.

Furthermore, the invention pertains to any novel chemical compounddescribed herein. That is, the invention relates to novel compounds, andnovel methods of their use as described herein, which are within thescope of the Formulae disclosed herein, and which are not disclosed inthe cited Patents and Patent Applications.

Synthesis of Compounds of the Invention

In general, the compounds of the present invention may be prepared bythe methods illustrated in the general reaction schemes as, for example,described below, or by modifications thereof, using readily availablestarting materials, reagents and conventional synthesis procedures. Inthese reactions, it is also possible to make use of variants which arein themselves known, but are not mentioned here. Functional andstructural equivalents of the compounds described herein and which havethe same general properties, wherein one or more simple variations ofsubstituents are made which do not adversely affect the essential natureor the utility of the compound are also included.

The compounds of the present invention may be readily prepared inaccordance with the synthesis schemes and protocols described herein, asillustrated in the specific procedures provided. However, those skilledin the art will recognize that other synthetic pathways for forming thecompounds of this invention may be used, and that the following isprovided merely by way of example, and is not limiting to the presentinvention. See, e.g., “Comprehensive Organic Transformations” by R.Larock, VCH Publishers (1989). It will be further recognized thatvarious protecting and deprotecting strategies will be employed that arestandard in the art (See, e.g., “Protective Groups in Organic Synthesis”by Greene and Wuts). Those skilled in the relevant arts will recognizethat the selection of any particular protecting group (e.g., amine andcarboxyl protecting groups) will depend on the stability of theprotected moiety with regards to the subsequent reaction conditions andwill understand the appropriate selections.

Further illustrating the knowledge of those skilled in the art is thefollowing sampling of the extensive chemical literature: “Chemistry ofthe Amino Acids” by J. P. Greenstein and M. Winitz, John Wiley & Sons,Inc., New York (1961); “Comprehensive Organic Transformations” by R.Larock, VCH Publishers (1989); T. D. Ocain, et al., J. Med. Chem. 31,2193-99 (1988); E. M. Gordon, et al., J. Med. Chem. 31, 2199-10 (1988);“Practice of Peptide Synthesis” by M. Bodansky and A. Bodanszky,Springer-Verlag, New York (1984); “Protective Groups in OrganicSynthesis” by T. Greene and P. Wuts (1991); “Asymmetric Synthesis:Construction of Chiral Molecules Using Amino Acids” by G. M. Coppola andH. F. Schuster, John Wiley & Sons, Inc., New York (1987); “The ChemicalSynthesis of Peptides” by J. Jones, Oxford University Press, New York(1991); and “Introduction of Peptide Chemistry” by P. D. Bailey, JohnWiley & Sons, Inc., New York (1992).

The synthesis of compounds of the invention is carried out in a solvent.Suitable solvents are liquids at ambient room temperature and pressureor remain in the liquid state under the temperature and pressureconditions used in the reaction. Useful solvents are not particularlyrestricted provided that they do not interfere with the reaction itself(that is, they preferably are inert solvents), and they dissolve acertain amount of the reactants. Depending on the circumstances,solvents may be distilled or degassed. Solvents may be, for example,aliphatic hydrocarbons (e.g., hexanes, heptanes, ligroin, petroleumether, cyclohexane, or methylcyclohexane) and halogenated hydrocarbons(e.g., methylenechloride, chloroform, carbontetrachloride,dichloroethane, chlorobenzene, or dichlororbenzene); aromatichydrocarbons (e.g., benzene, toluene, tetrahydronaphthalene,ethylbenzene, or xylene); ethers (e.g., diglyme, methyl-tert-butylether, methyl-tert-amyl ether, ethyl-tert-butyl ether, diethylether,diisopropylether, tetrahydrofuran or methyltetrahydrofurans, dioxane,dimethoxyethane, or diethyleneglycol dimethylether); nitriles (e.g.,acetonitrile); ketones (e.g., acetone); esters (e.g., methyl acetate orethyl acetate); and mixtures thereof.

In general, after completion of the reaction, the product is isolatedfrom the reaction mixture according to standard techniques. For example,the solvent is removed by evaporation or filtration if the product issolid, optionally under reduced pressure. After the completion of thereaction, water may be added to the residue to make the aqueous layeracidic or basic and the precipitated compound filtered, although careshould be exercised when handling water-sensitive compounds. Similarly,water may be added to the reaction mixture with a hydrophobic solvent toextract the target compound. The organic layer may be washed with water,dried over anhydrous magnesium sulphate or sodium sulphate, and thesolvent is evaporated to obtain the target compound. The target compoundthus obtained may be purified, if necessary, e.g., by recrystallization,reprecipitation, chromatography, or by converting it to a salt byaddition of an acid or base.

The compounds of the invention may be supplied in a solution with anappropriate solvent or in a solvent-free form (e.g., lyophilized). Inanother aspect of the invention, the compounds and buffers necessary forcarrying out the methods of the invention may be packaged as a kit,optionally including a container. The kit may be commercially used fortreating or preventing amyloid-related disease according to the methodsdescribed herein and may include instructions for use in a method of theinvention. Additional kit components may include acids, bases, bufferingagents, inorganic salts, solvents, antioxidants, preservatives, or metalchelators. The additional kit components are present as purecompositions, or as aqueous or organic solutions that incorporate one ormore additional kit components. Any or all of the kit componentsoptionally further comprise buffers.

The term “container” includes any receptacle for holding the therapeuticcompound. For example, in one embodiment, the container is the packagingthat contains the compound. In other embodiments, the container is notthe packaging that contains the compound, i.e., the container is areceptacle, such as a box or vial that contains the packaged compound orunpackaged compound and the instructions for use of the compound.Moreover, packaging techniques are well known in the art. It should beunderstood that the instructions for use of the therapeutic compound maybe contained on the packaging containing the therapeutic compound, andas such the instructions form an increased functional relationship tothe packaged product.

Pharmaceutical Preparations

In another embodiment, the present invention relates to pharmaceuticalcompositions comprising agents according to any of the Formulae hereinfor the treatment of an amyloid-related disease, as well as methods ofmanufacturing such pharmaceutical compositions.

In general, the agents of the present invention may be prepared by themethods illustrated in the general reaction schemes as, for example, inthe patents and patent applications referred to herein, or bymodifications thereof, using readily available starting materials,reagents and conventional synthesis procedures. In these reactions, itis also possible to make use of variants which are in themselves known,but are not mentioned here. Functional and structural equivalents of theagents described herein and which have the same general properties,wherein one or more simple variations of substituents are made which donot adversely affect the essential nature or the utility of the agentare also included.

The agents of the invention may be supplied in a solution with anappropriate solvent or in a solvent-free form (e.g., lyophilized). Inanother aspect of the invention, the agents and buffers necessary forcarrying out the methods of the invention may be packaged as a kit. Thekit may be commercially used according to the methods described hereinand may include instructions for use in a method of the invention.Additional kit components may include acids, bases, buffering agents,inorganic salts, solvents, antioxidants, preservatives, or metalchelators. The additional kit components are present as purecompositions, or as aqueous or organic solutions that incorporate one ormore additional kit components. Any or all of the kit componentsoptionally further comprise buffers.

The therapeutic agent may also be administered parenterally,intraperitoneally, intraspinally, or intracerebrally. Dispersions can beprepared in glycerol, liquid polyethylene glycols, and mixtures thereofand in oils. Under ordinary conditions of storage and use, thesepreparations may contain a preservative to prevent the growth ofmicroorganisms.

To administer the therapeutic agent by other than parenteraladministration, it may be necessary to coat the agent with, orco-administer the agent with, a material to prevent its inactivation.For example, the therapeutic agent may be administered to a subject inan appropriate carrier, for example, liposomes, or a diluent.Pharmaceutically acceptable diluents include saline and aqueous buffersolutions. Liposomes include water-in-oil-in-water CGF emulsions as wellas conventional liposomes (Strejan et al., J. Neuroimmunol. 7, 27(1984)).

Pharmaceutical compositions suitable for injectable use include sterileaqueous solutions (where water soluble) or dispersions and sterilepowders for the extemporaneous preparation of sterile injectablesolutions or dispersion. In all cases, the composition must be sterileand must be fluid to the extent that easy syringability exists. It mustbe stable under the conditions of manufacture and storage and must bepreserved against the contaminating action of microorganisms such asbacteria and fungi.

Suitable pharmaceutically acceptable vehicles include, withoutlimitation, any non-immunogenic pharmaceutical adjuvants suitable fororal, parenteral, nasal, mucosal, transdermal, intravascular (IV),intraarterial (IA), intramuscular (IM), and subcutaneous (SC)administration routes, such as phosphate buffer saline (PBS).

The vehicle can be a solvent or dispersion medium containing, forexample, water, ethanol, polyol (for example, glycerol, propyleneglycol, and liquid polyethylene glycol, and the like), suitable mixturesthereof, and vegetable oils. The proper fluidity can be maintained, forexample, by the use of a coating such as lecithin, by the maintenance ofthe required particle size in the case of dispersion and by the use ofsurfactants. Prevention of the action of microorganisms can be achievedby various antibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In manycases, isotonic agents are included, for example, sugars, sodiumchloride, or polyalcohols such as mannitol and sorbitol, in thecomposition. Prolonged absorption of the injectable compositions can bebrought about by including in the composition an agent which delaysabsorption, for example, aluminum monostearate or gelatin.

Sterile injectable solutions can be prepared by incorporating thetherapeutic agent in the required amount in an appropriate solvent withone or a combination of ingredients enumerated above, as required,followed by filtered sterilization. Generally, dispersions are preparedby incorporating the therapeutic agent into a sterile vehicle whichcontains a basic dispersion medium and the required other ingredientsfrom those enumerated above. In the case of sterile powders for thepreparation of sterile injectable solutions, the methods of preparationare vacuum drying and freeze-drying which yields a powder of the activeingredient (i.e., the therapeutic agent) plus any additional desiredingredient from a previously sterile-filtered solution thereof.

The therapeutic agent can be orally administered, for example, with aninert diluent or an assimilable edible carrier. The therapeutic agentand other ingredients may also be enclosed in a hard or soft shellgelatin capsule, compressed into tablets, or incorporated directly intothe subject's diet. For oral therapeutic administration, the therapeuticagent may be incorporated with excipients and used in the form ofingestible tablets, buccal tablets, troches, capsules, elixirs,suspensions, syrups, wafers, and the like. The percentage of thetherapeutic agent in the compositions and preparations may, of course,be varied. The amount of the therapeutic agent in such therapeuticallyuseful compositions is such that a suitable dosage will be obtained.

It is especially advantageous to formulate parenteral compositions indosage unit form for ease of administration and uniformity of dosage.Dosage unit form as used herein refers to physically discrete unitssuited as unitary dosages for the subjects to be treated; each unitcontaining a predetermined quantity of therapeutic agent calculated toproduce the desired therapeutic effect in association with the requiredpharmaceutical vehicle. The specification for the dosage unit forms ofthe invention are dictated by and directly dependent on (a) the uniquecharacteristics of the therapeutic agent and the particular therapeuticeffect to be achieved, and (b) the limitations inherent in the art ofcompounding such a therapeutic agent for the treatment of amyloiddeposition in subjects.

The present invention therefore includes pharmaceutical formulationscomprising the agents of the Formulae described herein, includingpharmaceutically acceptable salts thereof, in pharmaceuticallyacceptable vehicles for aerosol, oral and parenteral administration.Also, the present invention includes such agents, or salts thereof,which have been lyophilized and which may be reconstituted to formpharmaceutically acceptable formulations for administration, as byintravenous, intramuscular, or subcutaneous injection. Administrationmay also be intradermal or transdermal.

In accordance with the present invention, an agent of the Formulaedescribed herein, and pharmaceutically acceptable salts thereof, may beadministered orally or through inhalation as a solid, or may beadministered intramuscularly or intravenously as a solution, suspensionor emulsion. Alternatively, the agents or salts may also be administeredby inhalation, intravenously or intramuscularly as a liposomalsuspension.

Pharmaceutical formulations are also provided which are suitable foradministration as an aerosol, by inhalation. These formulations comprisea solution or suspension of the desired agent of any Formula herein, ora salt thereof, or a plurality of solid particles of the agent or salt.The desired formulation may be placed in a small chamber and nebulized.Nebulization may be accomplished by compressed air or by ultrasonicenergy to form a plurality of liquid droplets or solid particlescomprising the agents or salts. The liquid droplets or solid particlesshould have a particle size in the range of about 0.5 to about 5microns. The solid particles can be obtained by processing the solidagent of any Formula described herein, or a salt thereof, in anyappropriate manner known in the art, such as by micronization. The sizeof the solid particles or droplets will be, for example, from about 1 toabout 2 microns. In this respect, commercial nebulizers are available toachieve this purpose.

A pharmaceutical formulation suitable for administration as an aerosolmay be in the form of a liquid, the formulation will comprise awater-soluble agent of any Formula described herein, or a salt thereof,in a carrier which comprises water. A surfactant may be present whichlowers the surface tension of the formulation sufficiently to result inthe formation of droplets within the desired size range when subjectedto nebulization.

Peroral compositions also include liquid solutions, emulsions,suspensions, and the like. The pharmaceutically acceptable vehiclessuitable for preparation of such compositions are well known in the art.Typical components of carriers for syrups, elixirs, emulsions andsuspensions include ethanol, glycerol, propylene glycol, polyethyleneglycol, liquid sucrose, sorbitol and water. For a suspension, typicalsuspending agents include methyl cellulose, sodium carboxymethylcellulose, tragacanth, and sodium alginate; typical wetting agentsinclude lecithin and polysorbate 80; and typical preservatives includemethyl paraben and sodium benzoate. Peroral liquid compositions may alsocontain one or more components such as sweeteners, flavoring agents andcolorants disclosed above.

Pharmaceutical compositions may also be coated by conventional methods,typically with pH or time-dependent coatings, such that the subjectagent is released in the gastrointestinal tract in the vicinity of thedesired topical application, or at various times to extend the desiredaction. Such dosage forms typically include, but are not limited to, oneor more of cellulose acetate phthalate, polyvinylacetate phthalate,hydroxypropyl methyl cellulose phthalate, ethyl cellulose, waxes, andshellac.

Other compositions useful for attaining systemic delivery of the subjectagents include sublingual, buccal and nasal dosage forms. Suchcompositions typically comprise one or more of soluble filler substancessuch as sucrose, sorbitol and mannitol; and binders such as acacia,microcrystalline cellulose, carboxymethyl cellulose and hydroxypropylmethyl cellulose. Glidants, lubricants, sweeteners, colorants,antioxidants and flavoring agents disclosed above may also be included.

The compositions of this invention can also be administered topically toa subject, e.g., by the direct laying on or spreading of the compositionon the epidermal or epithelial tissue of the subject, or transdermallyvia a “patch”. Such compositions include, for example, lotions, creams,solutions, gels and solids. These topical compositions may comprise aneffective amount, usually at least about 0.1%, or even from about 1% toabout 5%, of an agent of the invention. Suitable carriers for topicaladministration typically remain in place on the skin as a continuousfilm, and resist being removed by perspiration or immersion in water.Generally, the carrier is organic in nature and capable of havingdispersed or dissolved therein the therapeutic agent. The carrier mayinclude pharmaceutically acceptable emolients, emulsifiers, thickeningagents, solvents and the like.

In one embodiment, active agents are administered at a therapeuticallyeffective dosage sufficient to inhibit amyloid deposition in a subject.A “therapeutically effective” dosage inhibits amyloid deposition by, forexample, at least about 20%, or by at least about 40%, or even by atleast about 60%, or by at least about 80% relative to untreatedsubjects. In the case of an Alzheimer's subject, a “therapeuticallyeffective” dosage stabilizes cognitive function or prevents a furtherdecrease in cognitive function (i.e., preventing, slowing, or stoppingdisease progression). The present invention accordingly providestherapeutic drugs. By “therapeutic” or “drug” is meant an agent having abeneficial ameliorative or prophylactic effect on a specific disease orcondition in a living human or non-human animal.

In the case of AA or AL amyloidosis, the agent may improve or stabilizespecific organ function. As an example, renal function may be stabilizedor improved by 10% or greater, 20% or greater, 30% or greater, 40% orgreater, 50% or greater, 60% or greater, 70% or greater, 80% or greater,or by greater than 90%.

In the case of IAPP, the agent may maintain or increase β-islet cellfunction, as determined by insulin concentration or the Pro-IAPP/IAPPratio. In a further embodiment, the Pro-IAPP/IAPP ratio is increased byabout 10% or greater, about 20% or greater, about 30% or greater, about40% or greater, or by about 50%. In a further embodiment, the ratio isincreased up to 50%. In addition, a therapeutically effective amount ofthe agent may be effective to improve glycemia or insulin levels.

In another embodiment, the active agents are administered at atherapeutically effective dosage sufficient to treat AA (secondary)amyloidosis and/or AL (primary) amyloidosis, by stabilizing renalfunction, decreasing proteinuria, increasing creatinine clearance (e.g.,by at least 50% or greater or by at least 100% or greater), remission ofchronic diarrhea, or by weight gain (e.g., 10% or greater). In addition,the agents may be administered at a therapeutically effective dosagesufficient to improve nephrotic syndrome.

Furthermore, active agents may be administered at a therapeuticallyeffective dosage sufficient to decrease deposition in a subject ofamyloid protein, e.g., Aβ40 or Aβ42. A therapeutically effective dosagedecreases amyloid deposition by, for example, at least about 15%, or byat least about 40%, or even by at least 60%, or at least by about 80%relative to untreated subjects.

In another embodiment, active agents are administered at atherapeutically effective dosage sufficient to increase or enhanceamyloid protein, e.g., Aβ40 or Aβ42, in the blood, CSF, or plasma of asubject. A therapeutically effective dosage increases the concentrationby, for example, at least about 15%, or by at least about 40%, or evenby at least 60%, or at least by about 80% relative to untreatedsubjects.

In yet another embodiment, active agents are administered at atherapeutically effective dosage sufficient to maintain a subject's CDRrating at its base line rating or at 0. In another embodiment, theactive agents are administered at a therapeutically effective dosagesufficient to decrease a subject's CDR rating by about 0.25 or more,about 0.5 or more, about 1.0 or more, about 1.5 or more, about 2.0 ormore, about 2.5 or more, or about 3.0 or more. In another embodiment,the active agents are administered at a therapeutically effective dosagesufficient to reduce the rate of the increase of a subject's CDR ratingas compared to historical or untreated controls. In another embodiment,the therapeutically effective dosage is sufficient to reduce the rate ofincrease of a subject's CDR rating (relative to untreated subjects) byabout 5% or greater, about 10% or greater, about 20% or greater, about25% or greater, about 30% or greater, about 40% or greater, about 50% orgreater, about 60% or greater, about 70% or greater, about 80% orgreater, about 90% or greater or about 100% or greater.

In yet another embodiment, active agents are administered at atherapeutically effective dosage sufficient to maintain a subject'sscore on the MMSE. In another embodiment, the active agents areadministered at a therapeutically effective dosage sufficient toincrease a subject's MMSE score by about 1, about 2, about 3, about 4,about 5, about 7.5, about 10, about 12.5, about 15, about 17.5, about20, or about 25 points. In another embodiment, the active agents areadministered at a therapeutically effective dosage sufficient to reducethe rate of the decrease of a subject's MMSE score as compared tohistorical controls. In another embodiment, the therapeuticallyeffective dosage is sufficient to reduce the rate of decrease of asubject's MMSE score may be about 5% or less, about 10% or less, about20% or less, about 25% or less, about 30% or less, about 40% or less,about 50% or less, about 60% or less, about 70% or less, about 80% orless, about 90% or less or about 100% or less, of the decrease of thehistorical or untreated controls.

In yet another embodiment, active agents are administered at atherapeutically effective dosage sufficient to maintain a subject'sscore on the ADAS-Cog. In another embodiment, the active agents areadministered at a therapeutically effective dosage sufficient todecrease a subject's ADAS-Cog score by about 2 points or greater, byabout 3 points or greater, by about 4 points or greater, by about 5points or greater, by about 7.5 points or greater, by about 10 points orgreater, by about 12.5 points or greater, by about 15 points or greater,by about 17.5 points or greater, by about 20 points or greater, or byabout 25 points or greater. In another embodiment, the active agents areadministered at a therapeutically effective dosage sufficient to reducethe rate of the increase of a subject's ADAS-Cog scores as compared tohistorical or untreated controls. In another embodiment, thetherapeutically effective dosage is sufficient to reduce the rate ofincrease of a subject's ADAS-Cog scores (relative to untreated subjects)by about 5% or greater, about 10% or greater, about 20% or greater,about 25% or greater, about 30% or greater, about 40% or greater, about50% or greater, about 60% or greater, about 70% or greater, about 80% orgreater, about 90% or greater or about 100% or greater.

In another embodiment, active agents are administered at atherapeutically effective dosage sufficient to decrease the ratio ofAβ42:Aβ40 in the CSF or plasma of a subject by about 15% or more, about20% or more, about 25% or more, about 30% or more, about 35% or more,about 40% or more, about 45% or more, or about 50% or more.

In another embodiment, active agents are administered at atherapeutically effective dosage sufficient to lower levels of Aβ in theCSF or plasma of a subject by about 15% or more, about 25% or more,about 35% or more, about 45% or more, about 55% or more, about 75% ormore, or about 95% or more.

Toxicity and therapeutic efficacy of such agents can be determined bystandard pharmaceutical procedures in cell cultures or experimentalanimals, e.g., for determining the LD50 (the dose lethal to 50% of thepopulation) and the ED50 (the dose therapeutically effective in 50% ofthe population). The dose ratio between toxic and therapeutic effects isthe therapeutic index and can be expressed as the ratio LD50/ED50, andusually a larger therapeutic index is more efficacious. While agentsthat exhibit toxic side effects may be used, care should be taken todesign a delivery system that targets such agents to the site ofaffected tissue in order to minimize potential damage to unaffectedcells and, thereby, reduce side effects.

It is understood that appropriate doses depend upon a number of factorswithin the ken of the ordinarily skilled physician, veterinarian, orresearcher. The dose(s) of the small molecule will vary, for example,depending upon the identity, size, and condition of the subject orsample being treated, further depending upon the route by which thecomposition is to be administered, if applicable, and the effect whichthe practitioner desires the small molecule to have upon the subject.Exemplary doses include milligram or microgram amounts of the smallmolecule per kilogram of subject or sample weight (e.g., about 1microgram per kilogram to about 500 milligrams per kilogram, about 100micrograms per kilogram to about 5 milligrams per kilogram, or about 1microgram per kilogram to about 50 micrograms per kilogram). It isfurthermore understood that appropriate doses depend upon the potency.Such appropriate doses may be determined using the assays describedherein. When one or more of these compounds is to be administered to ananimal (e.g., a human), a physician, veterinarian, or researcher may,for example, prescribe a relatively low dose at first, subsequentlyincreasing the dose until an appropriate response is obtained. Inaddition, it is understood that the specific dose level for anyparticular animal subject will depend upon a variety of factorsincluding the activity of the specific agent employed, the age, bodyweight, general health, gender, and diet of the subject, the time ofadministration, the route of administration, the rate of excretion, andany drug combination.

The ability of an agent to inhibit amyloid deposition can be evaluatedin an animal model system that may be predictive of efficacy ininhibiting amyloid deposition in human diseases, such as a transgenicmouse expressing human APP or other relevant animal models where Aβdeposition is seen or for example in an animal model of AA amyloidosis.Likewise, the ability of an agent to prevent or reduce cognitiveimpairment in a model system may be indicative of efficacy in humans.Alternatively, the ability of an agent can be evaluated by examining theability of the agent to inhibit amyloid fibril formation in vitro, e.g.,using a fibrillogenesis assay such as that described herein, including aThT, CD, or EM assay. Also the binding of an agent to amyloid fibrilsmay be measured using a MS assay as described herein. The ability of theagent to protect cells from amyloid induced toxicity is determined invitro using biochemical assays to determine percent cell death inducedby amyloid protein. The ability of an agent to modulate renal functionmay also be evaluated in an appropriate animal model system.

The therapeutic agent of the invention may be also be administered exvivo to inhibit amyloid deposition or treat certain amyloid-relateddiseases, such as β₂M amyloidosis and other amyloidoses related todialysis. Ex vivo administration of the therapeutic agents of theinvention can be accomplished by contacting a body fluid (e.g., blood,plasma, etc.) with a therapeutic compound of the invention such that thetherapeutic compound is capable of performing its intended function andadministering the body fluid to the subject. The therapeutic compound ofthe invention may perform its function ex vivo (e.g., dialysis filter),in vivo (e.g., administered with the body fluid), or both. For example,a therapeutic compound of the invention may be used to reduce plasma β₂Mlevels and/or maintain β₂M in its soluble form ex vivo, in vivo, orboth.

Prodrugs

The present invention is also related to prodrugs of the agents of theFormulae disclosed herein. Prodrugs are agents which are converted invivo to active forms (see, e.g., R. B. Silverman, 1992, “The OrganicChemistry of Drug Design and Drug Action,” Academic Press, Chp. 8).Prodrugs can be used to alter the biodistribution (e.g., to allow agentswhich would not typically enter the reactive site of the protease) orthe pharmacokinetics for a particular agent. For example, a carboxylicacid group, can be esterified, e.g., with a methyl group or an ethylgroup to yield an ester. When the ester is administered to a subject,the ester is cleaved, enzymatically or non-enzymatically, reductively,oxidatively, or hydrolytically, to reveal the anionic group. An anionicgroup can be esterified with moieties (e.g., acyloxymethyl esters) whichare cleaved to reveal an intermediate agent which subsequentlydecomposes to yield the active agent. The prodrug moieties may bemetabolized in vivo by esterases or by other mechanisms to carboxylicacids.

Examples of prodrugs and their uses are well known in the art (see,e.g., Berge, et al., “Pharmaceutical Salts”, J. Pharm. Sci. 66, 1-19(1977)). The prodrugs can be prepared in situ during the final isolationand purification of the agents, or by separately reacting the purifiedagent in its free acid form with a suitable derivatizing agent.Carboxylic acids can be converted into esters via treatment with analcohol in the presence of a catalyst.

Examples of cleavable carboxylic acid prodrug moieties includesubstituted and unsubstituted, branched or unbranched lower alkyl estermoieties, (e.g., ethyl esters, propyl esters, butyl esters, pentylesters, cyclopentyl esters, hexyl esters, cyclohexyl esters), loweralkenyl esters, dilower alkyl-amino lower-alkyl esters (e.g.,dimethylaminoethyl ester), acylamino lower alkyl esters, acyloxy loweralkyl esters (e.g., pivaloyloxymethyl ester), aryl esters (phenylester), aryl-lower alkyl esters (e.g., benzyl ester), substituted (e.g.,with methyl, halo, or methoxy substituents) aryl and aryl-lower alkylesters, amides, lower-alkyl amides, dilower alkyl amides, and hydroxyamides.

Pharmaceutically Acceptable Salts

Certain embodiments of the present agents can contain a basic functionalgroup, such as amino or alkylamino, and are, thus, capable of formingpharmaceutically acceptable salts with pharmaceutically acceptableacids. The term “pharmaceutically acceptable salts” in this respect,refers to the relatively non-toxic, inorganic and organic acid additionsalts of agents of the present invention. These salts can be prepared insitu during the final isolation and purification of the agents of theinvention, or by separately reacting a purified agent of the inventionin its free base form with a suitable organic or inorganic acid, andisolating the salt thus formed.

Representative salts include the hydrohalide (including hydrobromide andhydrochloride), sulfate, bisulfate, phosphate, nitrate, acetate,valerate, oleate, palmitate, stearate, laurate, benzoate, lactate,phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate,napthylate, mesylate, glucoheptonate, lactobionate,2-hydroxyethanesulfonate, and laurylsulphonate salts and the like. See,e.g., Berge et al., “Pharmaceutical Salts”, J. Pharm. Sci. 66, 1-19(1977).

In other cases, the agents of the present invention may contain one ormore acidic functional groups and, thus, are capable of formingpharmaceutically acceptable salts with pharmaceutically acceptablebases. The term “pharmaceutically acceptable salts” in these instancesrefers to the relatively non-toxic, inorganic and organic base additionsalts of agents of the present invention.

These salts can likewise be prepared in situ during the final isolationand purification of the agents, or by separately reacting the purifiedagent in its free acid form with a suitable base, such as the hydroxide,carbonate or bicarbonate of a pharmaceutically acceptable metal cation,with ammonia, or with a pharmaceutically acceptable organic primary,secondary or tertiary amine. Representative alkali or alkaline earthsalts include the lithium, sodium, potassium, calcium, magnesium, andaluminum salts and the like. Representative organic amines useful forthe formation of base addition salts include ethylamine, diethylamine,ethylenediamine, ethanolamine, diethanolamine, piperazine and the like.

“Pharmaceutically acceptable salts” also includes, for example,derivatives of agents modified by making acid or base salts thereof, asdescribed further below and elsewhere in the present application.Examples of pharmaceutically acceptable salts include mineral or organicacid salts of basic residues such as amines; and alkali or organic saltsof acidic residues such as carboxylic acids. Pharmaceutically acceptablesalts include the conventional non-toxic salts or the quaternaryammonium salts of the parent agent formed, for example, from non-toxicinorganic or organic acids. Such conventional non-toxic salts includethose derived from inorganic acids such as hydrochloric, hydrobromic,sulfuric, sulfamic, phosphoric, and nitric acid; and the salts preparedfrom organic acids such as acetic, propionic, succinic, glycolic,stearic, lactic, malic, tartaric, citric, ascorbic, palmoic, maleic,hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic,2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, and isethionic acid. Pharmaceutically acceptablesalts may be synthesized from the parent agent which contains a basic oracidic moiety by conventional chemical methods. Generally, such saltsmay be prepared by reacting the free acid or base forms of these agentswith a stoichiometric amount of the appropriate base or acid in water orin an organic solvent, or in a mixture of the two.

All acid, salt, base, and other ionic and non-ionic forms of thecompounds described are included as compounds of the invention. Forexample, if a compound is shown as an acid herein, the salt forms of thecompound are also included. Likewise, if a compound is shown as a salt,the acid and/or basic forms are also included.

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, numerous equivalents to thespecific procedures, embodiments, claims, and examples described herein.Such equivalents are considered to be within the scope of this inventionand covered by the claims appended hereto. The contents of allreferences, issued patents, and published patent applications citedthroughout this application are hereby incorporated by reference. Theinvention is further illustrated by the following examples, which shouldnot be construed as further limiting.

EXAMPLES

The Examples set forth herein below provide exemplary syntheses ofcertain representative compounds of the invention. Also provided areexemplary methods for assaying the compounds of the invention for Aβbinding affinity, in vivo efficacy, and neuroprotective activity.

Example 1 Synthesis of Representative Compounds of the Invention

The present invention also relates to novel compounds and the synthesisthereof. Accordingly, the following examples are presented to illustratehow some of those compounds may be prepared.

The synthetic protocols of compounds C; D; E; F; G; H; I; J; K; L; M; N;P; Q; R; S; X; Y; Z; AA; AB; AC; AD; AE; AF; AG; AH; AJ; AK; AL; AM; AV;AW; AY; AZ; BA; BB; BW; BY; BZ; CE; CG; CH; CI; CJ; CK; CL; CO; CV; DD;DG; DH; DI; DJ; DK; DL; DM; DO; DP; DQ; DR; DS; DT; DU; DV; DW; DX; DY;DZ; EA; EB; EC; ED; EE; EF; EG; EH; EI; EJ; EK; EN; EO; EP; EQ; ER; ES;ET; EV; EW; FN; FO are described at pages 155 to 201 of co-owned PCTpublication No. WO 2004/113275, which is incorporated herein in itsentirety.

Preparation of3-{[(1S)-1-benzyl-2-(benzyloxy)-2-oxoethyl]amino}propane-1-sulfonic acid(Compound EL)

L-Phenylalanine benzylester hydrochloride (2.0 g, 6.9 mmol) was treatedwith a saturated aqueous solution of K₂CO₃ (50 mL) and EtOAc (3×50 mL)was added. The organic extracts were separated, combined, dried withNa₂SO₄, filtered, evaporated under reduced pressure and dried in vacuo.

To a solution of L-Phenylalanine benzylester (1.8 g, 6.8 mmol) in1,4-dioxane (10 mL) was added 1,3-propanesultone (708 mg, 6.5 mmol). Thesolution was stirred at reflux. After 1 hour, 20 mL of 1,4-dioxane wasadded to allow for good stirring. The reaction was stirred at reflux foran additional 1 hour. It was cooled to room temperature. The solid wascollected by filtration, washed with acetone (2×25 mL). The product wassuspended 80% Acetone/EtOH. The suspension was stirred at reflux for 30seconds. The solid was filtered and dried in the vacuum oven (50° C.)affording the title compound (1.14 g, 46%). ¹H NMR (DMSO, 500 MHz) δ ppm7.27 (m, 6H), 7.12 (m, 4H), 5.05 (dd, 2H, J=12.3 Hz), 4.49 (m, 1H). 3.29(m, 1H), 3.00 (m, 1H), 2.98 (m, 1H), 2.61 (t, 2H, J=6.5 Hz), 1.97 (m,2H). ¹³C (DMSO, 125 MHz) δ ppm 168.88, 135.21, 134.90, 130.00, 129.31,129.07, 128.01, 67.98, 60.50, 49.77, 46.72, 35.87, 22.43. [α]_(D)=+4.8°(c=0.00073 in water), ES-MS 378 (M+1).

Preparation of3-{[(1S)-1-(methoxycarbonyl)-2-methylpropyl]amino}-1-propanesulfonicacid (Compound FT)

L-valine methylester hydrochloride (5.0 g, 29.8 mmol) was treated with asaturated solution of K₂CO₃ (75 mL) and EtOAc (3×75 mL) was added. Theorganic extracts were separated, combined, dried with Na₂SO₄, filtered,evaporated under reduced pressure and dried in vacuo.

To a solution of L-valine methylester in THF (25 mL) was slowly added1,3-propanesultone (2.49 g, 19.9 mmol). The solution was stirred atreflux for 2 hours. The reaction was cooled to room temperature. Thesolid was collected by filtration, washed with acetone (2×30 mL). It wasdried in the vacuum oven (50° C.) affording the title compound (2.52 g,50%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.92 (m, 1H), 3.75 (s, 3H), 3.13 (t,2H, J=6.8 Hz), 2.88 (t, 2H, J=6.8 Hz), 2.24 (m, 1H), 2.06 (m, 2H)), 0.96(d, 3H, J=6.8 Hz), 0.88 (d, 3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm169.35, 65.85, 55.61, 48.14, 45.59, 29.48, 21.32, 18.25, 16.57.[α]_(D)=+9.6° (c=0.0014 in water), ES-MS 254 (M+1).

Preparation of 3-{[(1R)-1-cyclohexylethyl]amino}-1-propanesulfonic acid(Compound FU)

To a solution of (R)-(−)-cyclohexylethylamine (2.5 g, 19.7 mmol) intetrahydrofuran (25 mL) was slowly added 1,3-propanesultone (2.33 g,18.7 mmol). The solution was stirred at reflux for 2 hours. The reactionwas cooled to room temperature. The solid was collected by filtration,washed with acetone (2×25 mL) and dried in vacuo, affording the titlecompound (3.47 g, 74%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.09 (m, 3H), 2.88(t, 2H, J=7.3 Hz), 2.00 (m, 2H), 1.58 (m, 6H), 1.13 (m, 5H), 1.03 (m,3H). ¹³C (D₂O, 125 MHz) δ ppm 58.37, 48.17, 44.00, 39.84, 29.00, 26.01,25.82, 25.73, 25.47, 21.51, 11.79. [α]_(D)=+4.5° (c=0.0022 in water),ES-MS 250 (M−1).

Preparation of 3-{[(1S)-1-cyclohexylethyl]amino}-1-propanesulfonic acid(Compound FW)

To a solution of (S)-(+)-cyclohexylethylamine (5.0 g, 39.3 mmol) intetrahydrofuran (50 mL) was slowly added 1,3-propanesultone (4.66 g,37.4 mmol). The solution was stirred at reflux for 2 hours. The reactionwas cooled to room temperature. The solid was collected by filtrationand washed with acetone (2×25 mL). The solid was suspended in 80%acetone/EtOH (200 mL). The suspension was stirred at reflux for 30seconds before the solid was filtered and dried in vacuo, affording thetitle compound (6.13 g, 66%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.09 (m, 3H),2.88 (t, 2H, J=7.3 Hz), 2.00 (m, 2H), 1.55 (m, 6H), 1.13 (m, 5H), 1.03(m, 3H). ¹³C (D₂O, 125 MHz) δ ppm 59.37, 48.17, 44.00, 39.84, 29.00,26.01, 25.82, 25.73, 25.47, 21.51, 11.78. [α]_(D)=−2.8° (c=0.0014 inwater), ES-MS 250 (M−1).

Preparation of 3-[(4-tert-butylcyclohexyl)amino]-1-propanesulfonic acid(Compound FX)

To a solution of 4-tert-butylcyclohexylethylamine (mixture of cis andtrans isomers, 2.5 g, 16.1 mmol) in tetrahydrofuran (30 mL) was slowlyadded 1,3-propanesultone (1.84 g, 15.3 mmol). The solution was stirredat reflux for 2 hours. The reaction was cooled to room temperature. Thesolid was collected by filtration and washed with acetone (2×35 mL). Thesolid was suspended in 80% acetone/EtOH (200 mL). The suspension wasstirred at reflux for 30 seconds before the solid was filtered and driedin vacuo, affording the title compound (3.07 g, 72%). ¹H NMR (DMSO, 500MHz) δ ppm 3.21 (m, 0.5H), 3.04 (m, 2H), 2.89 (m, 1H), 2.67 (m, 0.5H),1.97 (m, 4H), 1.77 (m, 1H), 1.52 (m, 1H), 1.19 (m, 2H), 0.96 (m, 2H),0.81 (s, 9H). ¹³C (DMSO, 125 MHz) δ ppm 56.47, 53.62, 50.44, 49.77,47.56, 47.04, 46.28, 44.49, 32.98, 32.74, 29.61, 28.10, 28.03, 25.57,22.68, 22.48, 21.01. ES-MS 276 (M−1).

Preparation of3-{[(1S,2S)-2-(benzyloxy)cyclopentyl]amino}-1-propanesulfonic acid(Compound FY)

To a solution of (1S,2S)-2-benzyloxycyclopentylamine (1.0 g, 5.2 mmol)in tetrahydrofuran (12 mL) was slowly added 1,3-propanesultone (601 mg,5.0 mmol). The solution was stirred at reflux for 2 hours. The reactionwas cooled to room temperature. The solid was collected by filtration,washed with acetone (2×25 mL) and dried in vacuo, affording the titlecompound (1.36 g, 87%). ¹H NMR (D₂O, 500 MHz) δ ppm 7.32 (m, 5H), 4.53(d, 1H, J=11.2 Hz), 4.41 (d, 1H, J=11.2 Hz), 4.01 (m, 1H), 3.36 (m, 1H),3.00 (t, 2H, J=7.8 Hz), 2.80 (t, 2H, J=7.8 Hz), 2.00 (m, 4H), 1.64 (m,3H), 1.49 (m, 1H). ¹³C NMR (D₂O, 125 MHz) δ ppm 136.99, 129.06, 129.01,128.77, 81.78, 71.81, 63.88, 48.01, 45.33, 29.91, 27.43, 21.60, 20.93.[α]_(D)=+31.1° (c=0.0064 in water). ES-MS 314 (M+1).

Preparation of3-{[(1R,2R)-2-(benzyloxy)cyclopentyl]amino}-1-propanesulfonic acid(Compound FZ)

To a solution of (1R,2R)-2-benzyloxycyclopentylamine (1.0 g, 5.2 mmol)in tetrahydrofuran (12 mL) was slowly added 1,3-propanesultone (601 mg,5.0 mmol). The solution was stirred at reflux for 2 hours. The reactionwas cooled to room temperature. The solid was collected by filtrationand washed with acetone (2×15 mL). The product was suspended in EtOH andthe solvent was evaporated (to remove THF residue). The solid was driedin vacuo, affording the title compound (717 mg, 46%). ¹H NMR (D₂O, 500MHz) δ ppm 7.32 (m, 5H), 4.53 (d, 1H, J=11.2 Hz), 4.42 (d, 1H, J=11.2Hz), 4.02 (m, 1H), 3.36 (m, 1H), 3.01 (t, 2H, J=7.8 Hz), 2.81 (t, 2H,J=7.8 Hz), 2.01 (m, 4H), 1.65 (m, 3H), 1.49 (m, 1H). ¹³C NMR (D₂O, 125MHz) δ ppm 137.00, 129.07, 129.01, 128.77, 81.78, 71.81, 63.89, 48.02,45.34, 29.93, 27.43, 21.61, 20.94. [α]_(D)=−38.8° (c=0.00122 in water).ES-MS 314 (M+1).

Preparation of3-{[(1S)-1-benzyl-2-(cyclohexylamino)-2-oxoethyl]amino}-1-propanesulfonicacid (Compound GA)

To a solution of L-Phenylalanine cyclohexylamide (2.5 g, 10.1 mmol) intetrahydrofuran (25 mL) was added 1,3-propanesultone (1.17 g, 9.7 mmol).The solution was stirred at reflux for 2 hours. It was cooled to roomtemperature. The solid was collected by filtration, washed with acetone(2×25 mL) and dried in the vacuum oven (50° C.), affording the titlecompound (1.39 g, 39%). ¹H NMR (D₂O, 500 MHz) δ ppm 7.21 (m, 3H), 7.08(m, 2H), 4.42 (m, 0.5H), 3.83 (m, 1H). 3.29 (m, 1H), 3.15 (m, 1H), 3.02(m, 2H), 2.86 (m, 3H), 2.49 (m, 0.5H), 2.01 (m, 2H), 1.54 (m, 1H), 1.45(m, 1H), 1.33 (m, 2H), 1.02 (m, 4H), 0.55 (m, 1H). ¹³C (D₂O, 125 MHz) δppm 133.71, 129.54, 129.18, 128.03, 62.08, 49.25, 47.97, 45.41, 36.29,31.73, 31.46, 24.86, 24.28, 24.20, 21.39. [α]_(D)=+36.4° (c=0.0019 inwater), ES-MS 369 (M+1).

Preparation of3-{[(1S,2S)-2-(benzyloxy)cyclopentyl]amino}-1-propanesulfonic acid(Compound GB)

To a solution of (1S,2S)-2-benzyloxycyclohexylamine (1.0 g, 5.2 mmol) intetrahydrofuran (12 mL) was slowly added 1,3-propanesultone (601 mg, 5.0mmol). The solution was stirred at reflux for 2 hours. The reaction wascooled to room temperature. The solid was collected by filtration,washed with acetone (2×20 mL) and dried in the vacuum oven (50° C.),affording the title compound (1.15 g, 75%). ¹H NMR (D₂O, 500 MHz) δ ppm7.34 (m, 5H), 4.62 (d, 1H, J=11.2 Hz), 4.42 (d, 1H, J=11.2 Hz), 3.40 (m,1H), 2.97 (m, 2H), 2.90 (m, 1H), 2.76 (t, 2H, J=6.5 Hz), 2.26 (m, 1H),1.92 (m, 3H), 1.66 (m, 2H), 1.18 (m, 4H). ¹³C NMR (D₂O, 125 MHz) δ ppm137.22, 129.34, 129.11, 128.84, 76.74, 70.26, 60.84, 48.02, 42.74,29.49, 26.43, 23.57, 23.02, 21.53. [α]_(D)=+74.8° (c=0.00207 in water).ES-MS 326 (M−1).

Preparation of3-{[(1R,2R)-2-(benzyloxy)cyclopentyl]amino}-1-propanesulfonic acid(Compound GD)

To a solution of (1R,2R)-2-benzyloxycyclohexylamine (1.0 g, 5.2 mmol) intetrahydrofuran (12 mL) was slowly added 1,3-propanesultone (601 mg, 5.0mmol). The solution was stirred at reflux for 2 hours. The reaction wascooled to room temperature. The solid was collected by filtration andwashed with acetone (2×35 mL). The solid was suspended in 80%acetone/EtOH (200 mL). The suspension was stirred at reflux for 30seconds before the solid was filtered and dried in the vacuum oven (50°C.), affording the title compound (844 mg. 55%). ¹H NMR (D₂O, 500 MHz) δppm 7.32 (m, 5H), 4.60 (d, 1H, J=11.2 Hz), 4.40 (d, 1H, J=11.2 Hz), 3.39(m, 1H), 2.94 (m, 2H), 2.85 (m, 1H), 2.74 (t, 2H, J=6.5 Hz), 2.24 (m,1H), 1.90 (m, 3H), 1.64 (m, 2H), 1.14 (m, 4H). ¹³C NMR (D₂O, 125 MHz) δppm 137.35, 129.28, 129.19, 128.90, 76.90, 70.35, 60.97, 48.12, 42.90,29.59, 26.53, 23.64, 23.09, 21.63. [α]_(D)=−68.9° (c=0.0026 in water).ES-MS 326 (M−1).

Preparation of3-({(1S)-1-[(benzyloxy)carbonyl]-2-methylpropyl}amino)-1-propanesulfonicacid (Compound GE)

L-valine benzylester p-tosylate (2.5 g, 6.6 mmol) was treated with asaturated solution of K₂CO₃ (50 mL) and EtOAc (3×50 mL). The organicextracts were separated, combined, dried with Na₂SO₄, filtered,evaporated under reduced pressure and dried in vacuo.

To a solution of L-valine benzylester in MeOH (12 mL) was slowly added1,3-propanesultone (604 mg, 5.0 mmol). The solution was stirred atreflux for 2 hours. The reaction was cooled to room temperature. Thesolid was collected by filtration, washed with cold MeOH and acetone(2×25 mL). It was dried in the vacuum oven (50° C.), affording the titlecompound (649 mg, 39%). ¹H NMR (D₂O, 500 MHz) δ ppm 7.47 (m, 5H), 5.41(d, 1H, J=11.7 Hz), 5.31 (d, 3H, J=11.7 Hz), 4.04 (m, 1H), 3.19 (m, 2H),2.95 (t, 2H, J=6.8 Hz), 2.35 (m, 1H), 2.14 (m, 2H), 1.04 (d, 3H, J=6.3Hz), 0.95 (d, 3H, J=6.3 Hz). ¹³C (D₂O, 125 MHz) δ ppm 168.75, 134.81,129.38, 129.31, 129.17, 69.00, 65.96, 48.24, 46.71, 29.67, 21.40, 18.39,16.65. [α]_(D)=−7.2° (c=0.0015 in water), ES-MS 330 (M+1).

Preparation of3-{[(1S)-2-ethoxy-1-methyl-2-oxoethyl]amino}-1-propanesulfonic acid(Compound GF)

L-alanine ethyl ester hydrochloride (2.5 g, 16.3 mmol) was treated witha saturated solution of K₂CO₃ (50 mL) and EtOAc (3×50 mL). The organicextracts were separated, combined, dried with Na₂SO₄, filtered,evaporated under reduced pressure and dried in vacuo.

To a solution of L-alanine ethyl ester (1.67 g, 14.3 mmol) intetrahydrofuran (25 mL) was slowly added 1,3-propanesultone (1.42 g,11.9 mmol). The solution was stirred at reflux for 2 hours. The reactionwas cooled to room temperature. The solid was collected by filtration,washed with acetone (2×25 mL) and dried in vacuo, affording the titlecompound (1.19 g, 42%). ¹H NMR (D₂O, 500 MHz) δ ppm 4.16 (m, 2H), 4.01(m, 1H), 3.12 (m, 2H), 2.87 (t, 2H, J=7.3 Hz), 2.01 (m, 2H), 1.43 (d,3H, J=7.3 Hz), 1.14 (t, 3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 170.22,63.84, 55.69, 47.94, 44.73, 21.51, 14.05, 13.25. [α]_(D)=−2.4° (c=0.0022in water), ES-MS 240 (M+1).

Preparation of (2S)-3-methyl-2-[(3-sulfopropyl)amino]butanoic acid(Compound GC)

L-valine methylester hydrochloride (5.0 g, 29.8 mmol) was treated with asaturated solution of K₂CO₃ (75 mL) and EtOAc (3×75 mL). The organicextracts were separated, combined, dried with Na₂SO₄, filtered,evaporated under reduced pressure and dried in vacuo.

To a solution of L-valine methylester in tetrahydrofuran (25 mL) wasslowly added 1,3-propanesultone (2.49 g, 19.9 mmol). The solution wasstirred at reflux for 2.5 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration, washed with acetone(2×30 mL) and dried in vacuo affording the desired ester.

The ester (860 mg, 3.4 mmol) was dissolved in 2M NaOH (1.20 g of NaOHand 15 mL of water). The reaction was stirred at room temperatureovernight. Dowex Marathon C ion exchange resin (strongly acidic) wasadded to the solution. The suspension was stirred for 15 minutes beforethe resin was removed by filtration. The filtrate was evaporated underreduced pressure and dried in vacuo, affording the title compound (645mg, 79%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.66 (m, 1H), 3.09 (t, 2H, J=6.3Hz), 2.86 (t, 2H, J=7.3 Hz), 2.17 (m, 1H), 2.07 (m, 2H)), 0.93 (d, 3H,J=6.8 Hz), 0.87 (d, 3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 171.24,66.83, 48.28, 46.77, 29.33, 21.44, 18.30, 16.98. [α]_(D)=−16.5°(c=0.0020 in water), ES-MS 238 (M−1).

Preparation of3-{[(1S)-1-(methoxycarbonyl)-3-methylbutyl]amino}-1-propanesulfonic acid(Compound GH)

L-Leucine methylester hydrochloride (5.0 g, 27.5 mmol) was treated witha saturated solution of K₂CO₃ (50 mL) and EtOAc (3×50 mL) was added. Theorganic extracts were separated, combined, dried with Na₂SO₄, filtered,evaporated under reduced pressure and dried in vacuo.

To a solution of L-valine methylester (3.74 g, 25.6 mmol) intetrahydrofuran (35 mL) was slowly added 1,3-propanesultone (2.04 g,17.2 mmol). The solution was stirred at reflux for 3 hours. The reactionwas cooled to room temperature. The solid was collected by filtration.Dowex Marathon C ion exchange resin (strongly acidic) was added to thesolution. The suspension was stirred for 15 minutes before the resin wasremoved by filtration. The filtrate was evaporated under reducedpressure. The solid was suspended in acetone (50 mL), filtered and driedin vacuo, affording the title compound (1.80 g, 39%). ¹H NMR (D₂O, 500MHz) δ ppm 3.99 (m, 1H), 3.72 (s, 3H), 3.12 (m, 2H), 2.87 (t, 2H, J=7.3Hz), 2.02 (m, 2H), 1.74 (m, 1H), 1.60 (m, 2H), 0.81 (d, 3H, J=5.4 Hz),0.87 (d, 3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 170.60, 58.91, 53.81,48.08, 45.50, 38.17, 24.44, 22.15, 21.59, 20.93. [α]_(D)=+13.8°(c=0.0016 in water), ES-MS 268 (M+1).

Preparation of3-({(1S)-1-[(tert-butylamino)carbonyl]-2-methylpropyl}amino)-1-propanesulfonicacid (Compound GI)

L-valine tert-butylamide hydrochloride (2.5 g, 12.0 mmol) was treatedwith a saturated solution of K₂CO₃ (50 mL) and EtOAc (3×50 mL) wasadded. The organic extracts were separated, combined, dried with Na₂SO₄,filtered, evaporated under reduced pressure and dried in vacuo.

To a solution of L-valine tert-butylamide (1.87 g, 11.0 mmol) in1,4-dioxane (20 mL) was added 1,3-propanesultone (1.07 g, 9.0 mmol). Thesolution was stirred at reflux for 5 hours. The reaction was cooled toroom temperature. The solid was collected by filtration, washed withacetone (2×30 mL) and dried in vacuo, affording the title compound (801mg, 25%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.43 (m, 1H), 3.00 (m, 2H), 2.85(m, 2H), 2.03 (m, 3H), 1.21 (m, 9H), 0.92 (d, 3H, J=6.3 Hz), 0.85 (d,3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 166.47, 66.55, 52.56, 48.23,46.11, 29.91, 27.81, 21.29, 18.30, 17.44. [α]_(D)=−11.6° (c=0.0023 inwater), ES-MS 293 (M−1).

Preparation of3-{[(1S)-1-(hydroxymethyl)-3-methylbutyl]amino}-1-propanesulfonic acid(Compound GJ)

To a solution of L-(+)-leucinol (5.0 g, 42.8 mmol) in THF (65 mL) wasslowly added 1,3-propanesultone (4.85 g, 40.7 mmol in THF (10 mL)). Thesolution was stirred at reflux for 2 hours. The reaction was cooled toroom temperature. The solid was collected by filtration and washed withacetone (2×50 mL). The solid was dissolved in 50% water/EtOH (400 mL).Dowex Marathon C ion exchange resin (strongly acidic) was added to thesolution. The suspension was stirred for 15 minutes before the resin wasremoved by filtration. The filtrate was evaporated under reducedpressure. The solid was suspended in acetone (150 mL), filtered anddried in the vacuum oven (50° C.), affording the title compound (6.11 g,63%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.77 (m, 1H), 3.59 (m, 1H), 3.23 (m,1H), 3.13 (m, 2H), 2.90 (m, 2H), 2.02 (m, 2H), 1.53 (m, 2H), 1.35 (m,1H), 0.81 (d, 3H, J=16.1 Hz). ¹³C (D₂O, 125 MHz) δ ppm 58.72, 58.00,48.17, 43.54, 35.96, 24.34, 22.53, 21.62, 20.89. [α]_(D)=+16.6°(c=0.0022 in water), ES-MS 240 (M+1).

Preparation of3-{[(1S)-1-(hydroxymethyl)-2-methylbutyl]amino}-1-propanesulfonic acid(Compound GK)

To a solution of L-(+)-isoleucinol (2.0 g, 17.1 mmol) in THF (30 mL) wasslowly added 1,3-propanesultone (1.94 g, 16.3 mmol). The solution wasstirred at reflux for 2 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration and washed withacetone (2×20 mL). The solid was dissolved in 70% water/EtOH (240 mL).Dowex Marathon C ion exchange resin (strongly acidic, 15 g) was added tothe solution. The suspension was stirred for 15 minutes before the resinwas removed by filtration. The filtrate was evaporated under reducedpressure. The solid was suspended in acetone (60 mL), filtered and driedin the vacuum oven (50° C.), affording the title compound (1.70 g, 44%).¹H NMR (D₂O, 500 MHz) δ ppm 3.78 (d, 1H, J=13.1 Hz), 3.64 (m, 1H), 3.14(m, 3H), 2.03 (m, 2H), 1.75 (m, 1H), 1.32 (m, 1H), 1.17 (m, 1H), 0.79(m, 6H). ¹³C (D₂O, 125 MHz) δ ppm 63.42, 57.38, 48.27, 44.77, 33.64,25.91, 21.52, 13.31, 10.94. [α]_(D)=+20.4° (c=0.00212 in water), ES-MS240 (M+1).

Preparation of3-{[(1R)-1-(hydroxymethyl)-3-methylbutyl]amino}-1-propanesulfonic acid(Compound GL)

To a solution of D-(−)-leucinol (2.0 g, 17.1 mmol) in THF (30 mL) wasslowly added 1,3-propanesultone (1.94 g, 16.3 mmol). The solution wasstirred at reflux for 2 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration and washed withacetone (2×20 mL). The solid was dissolved in 50% water/EtOH (240 mL).Dowex Marathon C ion exchange resin (strongly acidic) was added to thesolution. The suspension was stirred for 15 minutes before the resin wasremoved by filtration. The filtrate was evaporated under reducedpressure. The solid was suspended in acetone (50 mL), filtered and driedin the vacuum oven (50° C.), affording the title compound (2.55 g,(65%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.74 (d, 1H, J=12.7 Hz), 3.56 (d, 1H,J=12.7 Hz), 3.20 (m, 1H), 3.10 (t, 2H, J=7.3 Hz), 2.87 (t, 2H, J=7.3Hz), 2.00 (m, 2H), 1.49 (m, 2H), 1.31 (m, 1H), 0.80 (d, 3H, J=6.3 Hz),0.76 (d, 3H, J=6.3 Hz). ¹³C (D₂O, 125 MHz) δ ppm 58.74, 58.01, 48.19,43.56, 35.98, 24.34, 22.54, 21.63, 20.90. [α]_(D)=−16.3° (c=0.0019 inwater), ES-MS 238 (M−1).

Preparation of3-{[(1S)-2-amino-2-oxo-1-phenylethyl]amino}-1-propanesulfonic acid(Compound GN)

L-Phenylglycine amide hydrochloride (1.0 g, 6.7 mmol) was treated with asolution of K₂CO₃ (20 mL). The resultant mixture was extracted withEtOAc (3×20 mL). The organic extracts were separated, combined, driedover Na₂SO₄. The solid material was removed by filtration, and thefiltrate was concentrated to dryness under reduced pressure.

To a solution of L-Phenylglycine amide (670 mg, 5.9 mmol) intetrahydrofuran (10 mL) and 1,4-dioxane (4 mL) was added1,3-propanesultone (674 mg, 5.6 mmol). The solution stirred at refluxfor 2 hours. The reaction was cooled to room temperature. The solid wascollected by filtration and washed with acetone (2×20 mL). The solid wasdissolved in 50% water/EtOH mL). Dowex Marathon C ion exchange resin(strongly acidic) was added to the solution. The suspension was stirredfor 15 minutes before the resin was removed by filtration. The filtratewas evaporated under reduced pressure. The solid was suspended inacetone (50 mL), filtered and dried in the vacuum oven (50° C.),affording the title compound (743 mg, 50%). ¹H NMR (D₂O, 500 MHz) δ ppm7.38 (m, 5H), 4.92, (s, 1H), 3.01 (m, 1H), 2.91 (m, 1H), 2.78 (t, 2H,J=7.3 Hz), 2.0 (m, 2H). ¹³C (D₂O, 125 MHz) δ ppm 170.15, 130.95, 130.24,129.94, 128.74, 63.40, 47.99, 44.92, 21.27. [α]_(D)=−124° (c=0.0041 inwater), ES-MS 271 (M−1).

Preparation of3-{[(1S)-2-tert-butoxy-1-methyl-2-oxoethyl]amino}-1-propanesulfonic acid(Compound GO)

L-Alanine tert-butylester hydrochloride (2.61 g, 14.4 mmol) was treatedwith a solution of K₂CO₃ (75 mL). The resultant mixture was extractedwith EtOAc (3×75 mL). The organic extracts were separated, combined,dried over Na₂SO₄. The solid material was removed by filtration, and thefiltrate was concentrated to dryness under reduced pressure.

To a solution of L-Alanine tert-butylester (1.53 g, 10.5 mmol) intetrahydrofuran (20 mL) was added 1,3-propanesultone (1.16 g, 9.6 mmol).The solution stirred at reflux for 2 hours. The reaction was cooled toroom temperature. The solid was collected by filtration and washed withacetone (2×20 mL). The solid was dissolved in water (80 mL). DowexMarathon C ion exchange resin (strongly acidic) was added to thesolution. The suspension was stirred for 15 minutes before the resin wasremoved by filtration. The filtrate was evaporated under reducedpressure. The solid was suspended in acetone (80 mL), filtered and driedin the vacuum oven (50° C.), affording the title compound (1.37 g, 54%).¹H NMR (D₂O, 500 MHz) δ ppm 3.88 (m, 1H), 3.09 (m, 2H), 2.86 (t, 2H,J=7.3 Hz), 2.00 (m, 2H), 1.39 (d, 3H, J=7.3 Hz), 1.35 (s, 9H). ¹³C (D₂O,125 MHz) δ ppm 169.13, 86.12, 56.24, 47.94, 44.71, 27.11, 21.52, 14.17.[α]_(D)=−1.1° (c=0.0027 in water), ES-MS 266 (M−1).

Preparation of3-{[(1S)-2-amino-2-oxo-1-phenylethyl]amino}-1-propanesulfonic acid(Compound GP)

D-Phenylglycine amide hydrochloride (1.0 g, 6.7 mmol) was treated with asolution of K₂CO₃ (20 mL). The resultant mixture was extracted withEtOAc (3×20 mL). The organic extracts were separated, combined, driedover Na₂SO₄. The solid material was removed by filtration, and thefiltrate was concentrated to dryness under reduced pressure.

To a solution of D-Phenylglycine amide (850 mg, 7.5 mmol) intetrahydrofuran (10 mL) and 1,4-dioxane (4 mL) was added1,3-propanesultone (818 mg, 6.8 mmol). The solution stirred at refluxfor 2 hours. The reaction was cooled to room temperature. The solid wascollected by filtration and washed with acetone (2×20 mL). The solid wasdissolved in 50% water/EtOH mL). Dowex Marathon C ion exchange resin(strongly acidic) was added to the solution. The suspension was stirredfor 15 minutes before the resin was removed by filtration. The filtratewas evaporated under reduced pressure. The solid was suspended inacetone (50 mL), filtered and dried in the vacuum oven (50° C.),affording the title compound (720 mg, 34%). ¹H NMR (D₂O, 500 MHz) δ ppm7.38 (m, 5H), 4.92, (s, 1H), 3.00 (m, 1H), 2.90 (m, 1H), 2.78 (m, 2H),1.97 (m, 2H). ¹³C (D₂O, 125 MHz) δ ppm 170.14, 130.95, 130.24, 129.94,128.74, 63.40, 47.99, 44.92, 21.27. [α]_(D)=+106° (c=0.0016 in water),ES-MS 273 (M+1).

Preparation of (2S)-2-[(3-sulfopropyl)amino]propanoic acid (Compound GQ)

L-alanine methylester hydrochloride (5.0 g, 35.8 mmol) was treated witha saturated solution of K₂CO₃ (75 mL). The mixture was extracted withEtOAc (3×75 mL). The organic extracts were separated, combined, driedwith Na₂SO₄, filtered, evaporated under reduced pressure and dried invacuo.

To a solution of L-alanine methylester (2.37 g, 23.3 mmol) intetrahydrofuran (35 mL) was added 1,3-propanesultone (2.41 g, 20.0mmol). The solution was stirred at reflux for 2 hours. The reaction wascooled to room temperature. The solid was collected by filtration,washed with acetone (2×30 mL) and dried in vacuo.

The ester (2.21 g, 9.8 mmol) was dissolved in 2M NaOH (2.40 g of NaOHand 30 mL of water). The reaction was stirred at room temperatureovernight. Dowex Marathon C ion exchange resin (strongly acidic) wasadded to the solution. The suspension was stirred for 15 minutes beforethe resin was removed by filtration. The filtrate was evaporated underreduced pressure and lyophilized, affording the title compound (1.81 g,87%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.76 (m, 1H), 3.07 (m, 2H), 2.85 (t,2H, J=7.3 Hz), 1.99 (m, 2H), 1.38 (d, 3H, J=7.3 Hz), 0.87 (d, 3H, J=6.8Hz). ¹³C (D₂O, 125 MHz) δ ppm 173.31, 56.66, 47.97, 44.76, 21.56, 14.51.[α]_(D)=+3.5° (c=0.0023 in water), ES-MS 210 (M−1).

Preparation of (2S)-3-phenyl-2-[(3-sulfopropyl)amino]propanoic acid(Compound GR)

The N-(3-sulfo-propyl)-phenylalanine ethyl ester (DM-258-069, 860 mg,2.7 mmol) was dissolved in 2N NaOH (1.2 g of NaOH and 15 mL of water).The reaction was stirred at room temperature overnight. Dowex Marathon Cion exchange resin (strongly acidic) was added to the solution. Thesuspension was stirred for 15 minutes before the resin was removed byfiltration. The filtrate was evaporated under reduced pressure andlyophilized, affording the title compound (654 mg, 84%). ¹H NMR (D₂O,500 MHz) δ ppm 7.20 (m, 5H), 3.96 (t, 1H, J=6.3 Hz), 3.11 (m, 4H), 2.80(t, 2H, J=7.3 Hz), 1.95 (m, 2H). ¹³C (D₂O, 125 MHz) δ ppm 171.46,134.03, 129.50, 129.28, 128.10, 62.02, 47.97, 45.64, 35.23, 21.39.[α]_(D)32 +14.9° (c=0.0013 in water), ES-MS 286 (M−1).

Preparation of3-{[(1S)-1-isopropyl-2-oxopent-4-enyl]amino}-1-propanesulfonic acid(Compound GS)

L-Valine allylesterp-tosylate (3.0 g, 9.1 mmol) was treated with asaturated solution of K₂CO₃ (30 mL). The mixture was extracted withEtOAc (3×30 mL). The organic extracts were separated, combined, driedwith Na₂SO₄, filtered and evaporated under reduced pressure.

To a solution of L-valine allylester (1.30 g, 8.3 mmol) intetrahydrofuran (6 mL), 1,4-dioxane (6 mL) and MeOH (0.5 mL) was added1,3-propanesultone (910 mg, 7.5 mmol). The solution was stirred atreflux for 2 hours. The reaction was cooled to room temperature. Thesolvent was evaporated under reduced pressure. The sticky paste wassuspended in 20% acetone/ether. The solid was filtered and dissolvedEtOH (75 mL). Dowex Marathon C ion exchange resin (strongly acidic) wasadded to the solution. The suspension was stirred for 15 minutes beforethe resin was removed by filtration. The filtrate was evaporated todryness under reduced pressure, affording the title compound (605 mg,26%). ¹H NMR (D₂O, 500 MHz) δ ppm 5.84 (m, 1H), 5.27 (d, 1H, J=17.1 Hz),5.19 (m, 1H, J=10.3 Hz), 3.91 (d, 1H, J=3.9 Hz), 3.10 (t, 2H, J=7.3 Hz),2.85 (t, 2H, J=7.3 Hz), 2.22 (m, 1H), 2.03 (m, 2H), 0.93 (d, 3H, J=6.8Hz), 0.85 (d, 3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 168.55, 130.90,120.31, 67.58, 65.82, 48.09, 46.57, 29.53, 21.29, 18.25, 16.57.[α]_(D)=+5.0° (c=0.0011 in water), ES-MS 278 (M−1).

Preparation of3-{[(1S)-1-(aminocarbonyl)-3-methylbutyl]amino}-1-propanesulfonic acid(Compound GT)

L-Leucinamide hydrochloride (5.0 g, 30.0 mmol) was treated with asaturated solution of K₂CO₃ (100 mL). The mixture was extracted withEtOAc (3×100 mL). The organic extracts were separated, combined, driedwith Na₂SO₄, filtered and evaporated under reduced pressure.

To a solution of L-Leucinamide (3.20 g, 24.5 mmol) in tetrahydrofuran(35 mL) was added 1,3-propanesultone (2.82 g, 23.3 mmol). The solutionwas stirred at reflux for 2 hours. The reaction mixture was cooled toroom temperature. The solid was filtered and washed with acetone (2×25mL). The solid was dissolved in 50% EtOH/water (200 mL). Dowex MarathonC ion exchange resin (strongly acidic, 25 g) was added to the solution.The suspension was stirred for 15 minutes before the resin was removedby filtration. The filtrate was evaporated to dryness under reducedpressure. The solid was suspended in acetone (75 mL), and it was thenfiltered and dried in vacuo, affording the title compound (3.13 g, 53%).¹H NMR (D₂O, 500 MHz) δ ppm 3.79 (m, 1H), 3.04 (m, 2H), 2.85 (m, 2H),2.02 (m, 2H), 1.65 (m, 1H), 1.54 (m, 2H), 0.80 (m, 6H). ¹³C (D₂O, 125MHz) δ ppm 171.46, 59.42, 48.04, 45.46, 39.04, 24.27, 22.24, 21.49,21.17. [α]_(D)=+13.5° (c=0.0026 in water), ES-MS 251 (M−1).

Preparation of3-{[(1S)-1-(benzyloxycarbonyl)-3-methylbutyl]amino}-1-propanesulfonicacid (Compound GU)

L-Leucine benzylester p-tosylate (5.0 g, 12.7 mmol) was treated with asaturated solution of K₂CO₃ (100 mL). The mixture was extracted withEtOAc (3×100 mL). The organic extracts were separated, combined, driedwith Na₂SO₄, filtered and evaporated under reduced pressure.

To a solution of L-Leucine benzylester (2.81 g, 12.7 mmol) intetrahydrofuran (6 mL), 1,4-dioxane (6 mL) and MeOH (6 mL) was added1,3-propane sultone (1.40 g, 11.5 mmol). The solution was stirred atreflux for 2.5 hours. The reaction mixture was cooled to roomtemperature. The solid was filtered and washed with acetone (2×20 mL).The filtrate was evaporated under reduced pressure. The residue wasdissolved in acetone (20 mL). The product was precipitated with Et₂O(200 mL). The solid material was filtered. Both solids were combined anddissolved in 50% EtOH/water (200 mL). Dowex Marathon C ion exchangeresin (strongly acidic) was added to the solution. The suspension wasstirred for 15 minutes before the resin was removed by filtration. Thefiltrate was evaporated under reduced pressure and lyophilized,affording the title compound (1.87 g, 47%). ¹H NMR (DMSO, 500 MHz) δ ppm9.34 (s (broad), 1H), 7.39 (m, 5H), 5.25 (s, 2H), 4.10 (m, 1H), 3.09 (m,2H), 2.60 (m, 2H), 1.95 (m, 2H), 1.64 (m, 3H), 0.86 (m, 6H). ¹³C (DMSO,125 MHz) δ ppm 168.90, 134.91, 128.53, 128.50, 128.41, 67.38, 57.37,49.17, 45.79, 38.06, 24.07, 22.79, 21.78, 21.33. [α]_(D)=+1.8° (c=0.0017in water), ES-MS 344 (M+1).

Preparation of3-{[(1S)-1-(methyloxycarbonyl)-3-methylbutyl]amino}-1-propanesulfonicacid (Compound GZ)

L-Isoleucine methylester hydrochloride (5.0 g, 27.5 mmol) was treatedwith a saturated solution of K₂CO₃ (100 mL). The mixture was extractedwith EtOAc (3×100 mL). The organic layers were separated, combined,dried with Na₂SO₄, filtered and evaporated under reduced pressure.

To a solution of L-Isoleucine methlylester (3.43 g, 23.6 mmol) inacetone (30 mL) was added 1,3-propane sultone (2.62 g, 21.5 mmol). Thesolution was stirred at reflux for 2 h. The reaction mixture was cooledto room temperature. The solid was filtered and washed with acetone(2×20 mL). The filtrate was evaporated under reduced pressure. Theresidue was suspended in acetone (50 mL). The solid was filtered. Thesolid materials were combined and dissolved in water (100 mL). DowexMarathon C ion exchange resin (strongly acidic) was added to thesolution. The suspension was stirred for 15 minutes before the resin wasremoved by filtration. The filtrate was evaporated under reducedpressure. The solid product was suspended in acetone (100 mL), filteredand dried in vacuo, affording the title compound (3.23 g, 56%). ¹H NMR(D₂O, 500 MHz) δ ppm 4.00 (m, 1H), 3.74 (s, 3H), 3.14 (t, 2H, J=7.8 Hz),2.89 (t, 2H, J=7.3 Hz), 2.05 (m, 2H), 1.97 (m, 1H), 1.41 (m. 1H), 1.23(m, 1H), 0.83 (m, 6H). ¹³C (D₂O, 125 MHz) δ ppm 169.29, 64.51, 53.55,48.14, 46.52, 36.07, 25.92, 21.34, 13.76, 11.09. [α]_(D)=+22.6°(c=0.0023 in water), ES-MS 266 (M−1).

Preparation of3-{[(1S)-1-(oxycarbonyl)-3-methylbutyl]amino}-1-propanesulfonic acid(Compound HA)

L-Isoleucine methylester hydrochloride (5.0 g, 27.5 mmol) was treatedwith a saturated solution of K₂CO₃ (100 mL). The mixture was extractedwith EtOAc (3×100 mL). The organic layers were separated, combined,dried with Na₂SO₄, filtered and evaporated under reduced pressure.

To a solution of L-Isoleucine methlylester (3.43 g, 23.6 mmol) inacetone (30 mL) was added 1,3-propane sultone (2.62 g, 21.5 mmol). Thesolution was stirred at reflux for 2 hours. The reaction mixture wascooled to room temperature. The solid was filtered and washed withacetone (2×20 mL). The filtrate was evaporated under reduced pressure.The residue was suspended in acetone (50 mL). The solid was filtered.The solid materials were combined and dissolved in water (100 mL). DowexMarathon C ion exchange resin (strongly acidic) was added to thesolution. The suspension was stirred for 15 minutes before the resin wasremoved by filtration. The filtrate was evaporated under reducedpressure. The solid product was suspended in acetone (100 mL), filteredand dried in vacuo (3.23 g, 56%).

The solid (1.0 g, 3.7 mmol) was dissolved in 2M NaOH (30 mL). Thereaction mixture was stirred at room temperature overnight. DowexMarathon C ion exchange resin (strongly acidic, 15 g) was added to thesolution. The suspension was stirred for 15 minutes before the resin wasremoved by filtration. The filtrate was co-evaporated with EtOH andlyophilized, affording the title compound (740 mg, 83%). ¹H NMR (D₂O,500 MHz) δ ppm 4.00 (m, 1H), 3.59 (m, 3H), 3.07 (t, 2H, J=7.3 Hz), 2.86(m, 2H), 2.02 (m, 2H), 1.84 (m, 1H), 1.39 (m. 1H), 1.19 (m, 1H), 0.81(m, 6H). ¹³C (D₂O, 125 MHz) δ ppm 169.29, 64.51, 53.55, 48.14, 46.52,36.07, 25.92, 21.34, 13.76, 11.09. [α]_(D)=+30.4° (c=0.0031 in water),ES-MS 252 (M−1).

Preparation of3-{[(1S)-1-carbamoyl-2-phenylethyl]amino}-1-propanesulfonic acid(Compound HB)

L-Phenylalaninamide hydrochloride (5.0 g, 24.9 mmol) was treated with asaturated solution of K₂CO₃ (75 mL). The mixture was extracted withEtOAc (3×75 mL). The organic layers were separated, combined, dried withNa₂SO₄, filtered and evaporated under reduced pressure.

To a solution of L-Phenylalaninamide (3.93 g, 23.9 mmol) in acetonitrile(25 mL) was added 1,3-propane sultone (2.70 g, 21.8 mmol). The solutionwas stirred at reflux for 2 hours. The reaction mixture was cooled toroom temperature. The solid was filtered and washed with acetonitrile(2×25 mL). The solid product was suspended in EtOH (100 mL). Thesuspension was stirred at reflux for 1 hour. The solid material wasfiltered and dried in vacuo, affording the title compound (5.18 g, 83%).¹H NMR (D₂O, 500 MHz) δ ppm 7.24 (m, 3H), 7.16 (m, 2H), 4.02 (m, 1H),3.15 (m, 1H), 3.01 (m, 3H), 2.83 (m, 2H), 2.02 (m, 2H), 1.98 (m, 2H).¹³C (D₂O, 125 MHz) δ ppm 170.48, 133.72, 129.58, 129.21, 128.11, 61.55,47.95, 45.52, 36.21, 21.44. [α]_(D)=+23.1° (c=0.0021 in water), ES-MS285 (M−1).

Preparation of3-{[(1R)-1-(methoxycarbonyl)-3-methylbutyl]amino}-1-propanesulfonic acid(Compound HC)

D-Leucine methylester hydrochloride (2.63 g, 14.5 mmol) was treated witha saturated solution of K₂CO₃ (50 mL). The aqueous mixture was extractedwith EtOAc (3×50 mL). The organic layers were separated, combined, driedwith Na₂SO₄, filtered and evaporated under reduced pressure.

To a solution of D-Leucine methylester (1.58 g, 10.9 mmol) inacetonitrile (35 mL) was added 1,3-propanesultone (1.21 g, 9.9 mmol).The solution was stirred at reflux for 2 hours. The reaction mixture wascooled to room temperature. The solid material was collected byfiltration, recrystallized from EtOH and dried in vacuo, affording thetitle compound (1.59 g, 60%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.98 (m, 1H),3.70 (s, 3H), 3.11 (m, 2H), 2.85 (m, 2H), 2.00 (m, 2H), 1.72 (m, 1H),1.60 (m, 2H), 0.81 (m, 6H). ¹³C (D₂O, 125 MHz) δ ppm 170.51, 58.78,53.69, 47.97, 45.36, 38.09, 24.34, 22.07, 21.51, 20.82. [α]_(D)=+13.1°(c=0.0019 in water), ES-MS 266 (M−1).

Preparation of3-{[(1R)-1-(aminocarbonyl)-2-methylpropyl]amino}-1-propanesulfonic acid(Compound HD)

D-Valinamide hydrochloride (2.49 g, 14.7 mmol) was treated with asolution of K₂CO₃ (50 mL). The organic mixture was extracted with EtOAc(3×50 mL). The organic extracts were separated, combined, dried withNa₂SO₄, filtered, evaporated under reduced pressure and dried in vacuo.

To a solution of D-valinamide (1.76 g, 14.7 mmol) in acetonitrile (30mL) was slowly added 1,3-propanesultone (1.75 g, 14.4 mmol). Thesolution was stirred at reflux for 2 hours.

The reaction mixture was cooled to room temperature. The solid wascollected by filtration, washed with acetonitrile (2×25 mL). The solidproduct was dissolved in water (75 mL). Dowex Marathon C resin (stronglyacidic) was added to the solution. The suspension was stirred for 15minutes before the resin was removed by filtration. The filtrate wasevaporated under reduced pressure. The solid material was suspended inacetone (50 mL), filtered and dried in vacuo, affording the titlecompound (1.57 g, 51%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.80 (m, 1H), 3.19(m, 2H), 3.00 (m, 2H), 2.25 (m, 1H), 2.16 (m, 2H), 1.08 (d, 3H, J=6.8Hz), 1.02 (d, 3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 169.94, 65.86,48.10, 46.27, 29.54, 21.23, 17.99, 17.02. [α]_(D)=−12.4° (c=0.0037 inwater), ES-MS 237 (M−1).

Preparation of3-{[(1R)-1-carbamoyl-2-phenylethyl]amino}-1-propanesulfonic acid(Compound HE)

D-Phenylalaninamide hydrochloride (2.53 g, 12.6 mmol) was treated with asaturated solution of K₂CO₃ (50 mL). The mixture was extracted withEtOAc (3×50 mL). The organic layers were separated, combined, dried withNa₂SO₄, filtered and evaporated under reduced pressure.

To a solution of D-Phenylalaninamide (1.83 g, 11.1 mmol) in acetonitrile(20 mL) was added 1,3-propane sultone (1.29 g, 10.6 mmol). The solutionwas stirred at reflux for 2.5 hours. The reaction mixture was cooled toroom temperature. The solid was filtered and washed with acetonitrile(2×20 mL). The solid product was suspended in EtOH (75 mL). Thesuspension was stirred at reflux for 1 hours. The solid material wasfiltered, washed with acetone (1×25 mL) and dried in vacuo, affordingthe title compound (2.62 g, 89%). ¹H NMR (D₂O, 500 MHz) δ ppm 7.28 (m,3H), 7.19 (m, 2H), 4.05 (m, 1H), 3.19 (dd, 1H, J=5.3 Hz, 14.2 Hz), 3.04(m, 3H), 2.86 (t, 2H, J=5.8 Hz), 2.03 (m, 2H). ¹³C (D₂O, 125 MHz) ppm170.39, 133.73, 129.62, 129.26, 128.15, 61.57, 47.99, 45.57, 36.21,21.45. [α]_(D)=−20.7° (c=0.0038 in water), ES-MS 285 (M−1).

Preparation of3-({(1S)-1-[(benzyloxy)carbonyl]-2-methylbutyl}amino)-1-propanesulfonicacid (Compound HF)

L-Isoleucine benzylester p-tosylate (2.50 g, 6.4 mmol) was treated witha saturated solution of K₂CO₃ (30 mL). The mixture was extracted withEtOAc (3×30 mL). The organic extracts were separated, combined, driedwith Na₂SO₄, filtered and evaporated under reduced pressure.

To a solution of L-isoleucine benzylester (1.41 g, 6.4 mmol) inacetonitrile (12 mL) was added 1,3-propane sultone (706 mg, 5.8 mmol).The solution was stirred at reflux for 2 hours. The reaction mixture wascooled to room temperature. The solid was filtered and washed withacetone (2×20 mL). The solid material was dissolved in 50% EtOH/water(50 mL). Dowex Marathon C ion exchange resin (strongly acidic, 10 g) wasadded to the solution. The suspension was stirred for 15 minutes beforethe resin was removed by filtration. The filtrate was evaporated underreduced pressure and lyophilized affording the title compound (778 mg,39%). ¹H NMR (D₂O, 500 MHz) δ ppm 7.49 (m, 5H), 5.42 (d, 1H, J=11.7 Hz),5.31 (d, 1H, J=11.7 Hz), 3.24 (m, 2H), 2.98 (m, 2H), 2.13 (m, 3H), 1.46(m; 1H), 1.34 (m, 1H), 0.93 (m, 6H). ¹³C (D₂O, 125 MHz) δ ppm 168.53,134.69, 129.28, 129.22, 129.06, 68.86, 64.45, 48.12, 46.56, 36.21,25.97, 21.31, 13.73, 11.05. [α]_(D)=−1.5° (c=0.0031 in water), ES-MS 342(M−1).

Preparation of3-{[(1R)-1-(aminocarbonyl)-3-methylbutyl]amino}-1-propanesulfonic acid(Compound HG)

D-Leucinamide hydrochloride (1.0 g, 6.0 mmol) was treated with asaturated solution of K₂CO₃ (30 mL). The aqueous mixture was extractedwith EtOAc (3×30 mL). The organic extracts were separated, combined,dried with Na₂SO₄, filtered and evaporated under reduced pressure.

To a solution of D-Leucinamide (6.0 mmol) in acetonitrile (35 mL) wasadded 1,3-propanesultone (666 mg, 5.5 mmol). The solution was stirred atreflux for 2 hours. The reaction mixture was cooled to room temperature.The solid was filtered and washed with MeCN (2×20 mL). The solid wassuspended in EtOH (50 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature. The solid material wasfiltered, washed with acetone (1×20 mL) and dried in a vacuum oven (50°C.), affording the title compound (1.03 g, 74%): ¹H NMR (D₂O, 500 MHz) δppm 3.81 (m, 1H), 3.07 (m, 2H), 2.85 (t, 2H, J=7.3 Hz), 2.03 (m, 2H),1.68 (t, 1H, J=7.8 Hz), 1.58 (m, 2H), 0.83 (m, 6H). ¹³C (D₂O, 125 MHz) δppm 171.45, 59.39, 48.01, 45.41, 39.02, 24.24, 22.21, 21.47, 21.13.[α]_(D)=−13.7° (c=0.0019 in water), ES-MS 251 (M−1).

3-[(1-methylcyclopentyl)amino]-1-propanesulfonic acid (Compound FQ)

For the Ritter reaction, the flask was closed with a septum andconnected to a 20% NaOH scrubber. Sodium cyanide (powdered, 5.5 g, 112mmol) was added to acetic acid (30 mL) in one portion. The mixture wasstirred for 10 minutes at room temperature. A solution of sulfuric acid(16 mL) in acetic acid (15 mL) was added dropwise over a 20 minuteperiod. Then, a solution of 1-methyl-1-cyclopentanol (10 g, 99.8 mmol)in acetic acid (5 mL) was added dropwise over a 5 minute period. Themixture was stirred at room temperature for 22 hours then poured overice (approx. 100 g). The pH of the solution was adjusted to 9 with theaddition of 50% NaOH (about 135 g). The layers were separated and theaqueous layer was extracted with ether (1×40 mL). The combined organiclayers were washed with saturated sodium carbonate (1×10 mL) then driedover sodium sulfate. The ether was evaporated under reduced pressure toafford light brown oil (12.04 g, 94.7 mmol, 95%). The oil showed to be amixture of cis and trans formamide but what otherwise pure enough to beused as such. ¹H NMR (500 MHz, CDCl₃) δ [1.40 and 1.45 (s, 3H)],1.68-1.76 (m, 7H), 1.97-1.98 (m, 1H), [5.42 and 6.24 (br s, 1H)], [8.05(s) and 8.24 (d, J=12.2 Hz) for 1H]; ¹³C NMR (125 MHz, CDCl₃) δ 23.1,23.6, 25.5, 28.2, 39.5, 40.7, 60.7, 61.3, 160.7, 163.9

A solution of NaOH (25%, 80 mL) was added to the crude1-methyl-1-cyclopentylformamide (12.00 g, 94.7 mmol). The mixture washeated to reflux for 2.5 hours then cooled at room temperature. Somesodium chloride (20 g) was added to facilitate the phase separation. Thelayers were separated and the aqueous layer was extracted with toluene(1×15 mL). The combined organic layers were agitated. The addition ofisopropyl ether (2.5 mL, chloroform (1 g) and cyclohexane (6.5 g) didnot improved the separation of the solution. The combined organic layerswere washed with brine (1×10 mL) then dried over sodium sulfate andfiltered. The filtrate was used as such in the next step. ¹H NMR (500MHz, CDCl₃) δ 1.22 (s, 3H), 1.47-1.75 (4 m, 9H): ¹³C NMR (125 MHz,CDCl₃) δ 24.1, 29.6, 42.2, 58.4

A solution of 1,3-propanesultone (9.4 g, 75 mmol) in 2-butanone (35 mL)was added dropwise to a the crude solution of1-methyl-1-cyclopentylamine (mixture of solvent from previous step).

The mixture was heated to reflux for 20 hours then was cooled to roomtemperature. The solid was collected by suction-filtration and rinsedwith acetone (2×10 mL). The solid was dried overnight at 45° C. in thevacuum oven. The title compound was obtained as a fine white solid(16.26 g, 73.47 mmol, 74% overall yield). ¹H NMR (500 MHz, D₂O) δ 1.31(s, 3H), 1.591.6-1-85 (m, 8H), 2.03-2.06 (m, 2H), 2.96 (t, J=6.8 Hz,2H), 3.15 (t, J=7.6 Hz, 2H); ¹³C NRM (125 MHz, D₂O) δ 22.1, 22.5, 23.6,36.5, 41.4, 48.1, 66.8.1; ES-MS 220 (M−H)

3-[(1-methylcyclohexyl)amino]-1-propanesulfonic acid (Compound FR)

For the Ritter reaction, the flask was closed with a septum andconnected to a 20% NaOH scrubber. Potassium cyanide (powdered, 3.3 g, 50mmol) was added in portions to acetic acid (13 mL). The mixture wasstirred for 10 minutes at room temperature. A solution of sulfuric acid(7 mL) in acetic acid (7 mL) was added drop-wise over a 10 minuteperiod. Then, a solution of 1-methyl-1-cyclohexanol (5 g, 43.8 mmol) inacetic acid (4 mL) was added dropwise over a 5 minute period. Themixture was stirred at room temperature for 22 hours then poured overice (approx. 50 g). The pH of the solution was adjusted to 9 with theaddition of 50% NaOH (about 70 g). The layers were separated and theaqueous layer was extracted with ether (2×20 mL). The combined organiclayers were washed with saturated sodium carbonate (1×10 mL) then driedover sodium sulfate. The ether was evaporated under reduced pressure toafford a clear yellow oil (6.56 g, quantitative). The oil showed to be amixture of cis and trans formamide but what otherwise pure enough to beused as such. ¹H NMR (500 MHz, CDCl₃) δ 1.33-1.53 (3 m, 11H), 1.67 (brs, 1H), 1.99 (m, 1H), [5.16 and 6.09 (br s, 1H)], [8.11 (s) and 8.25 (d,J=12.2 Hz) for 1H]; ¹³C NMR (125 MHz, CDCl₃) δ 23.1, 23.6, 25.5, 28.2,39.5, 40.7, 60.7, 61.3, 160.7, 163.9

A solution of NaOH (20%, 40 mL) was added to the crude1-methyl-1-cyclohexylformamide (43.8 mmol). The mixture was heated toreflux for 3 hours then cooled at room temperature. Some sodium chloride(7.5 g) was added to facilitate the phase separation. The layers wereseparated and the aqueous layer was extracted with MTBK (1×10 mL). Thecombined organic layers were washed with brine (1×5 mL) the dried oversodium sulfate and filtered. The filtrate was used as such in the nextstep. ¹H NMR (500 MHz, CDCl₃) δ 1.08 (s, 3H), 1.33-1.51 (m, 10H); ¹³CNMR (125 MHz, CDCl₃) δ 22.8, 25.8, 29.6, 40.8, 48.6

A solution of 1,3-propanesultone (5.00 g, 40 mmol) in toluene (10 mL)was added dropwise to a the crude solution of 1-methyl-1-cyclohexylaminein MTBK (total volume 30 mL). The mixture was heated to reflux for 18hours then cooled to room temperature. The solid was collected bysuction filtration, rinsed with acetone (2×8 mL). The solid was driedovernight at 45° C. in the vacuum oven. The title compound was obtainedas a fine white solid (9.22 g). However, the proton NMR and the ES-MSwere not clean. The solid was suspended in methanol (45 mL) and thesuspension was warmed to reflux. Water (12 mL) was added dropwise untila clear yellow solution was obtained. The mixture was slowly cool toroom temperature with stirring. The solid was collected bysuction-filtration, rinsed with methanol (2×5 mL). Another crop wascollected from the filtrate. Both crops were dried overnight at 45° C.in the vacuum oven. The title compound was obtained as a fine whitesolid (6.82 g, 29.0 mmol, 66% overall yield). Both crops were identicaland were mixed for submitting the compound. ¹H NMR (500 MHz, D₂O) δ1.04-1.11 (m, 1H), 1.19 (s, 3H), 1.31 (q, J=12.2 Hz, 2H), 1.40 (qt,J=12.2 Hz, 2H), 1.46-1.62 (m, 2H), 1.63 (br d, J=11.7 Hz, 2H), 1.94 (q,J=7.3 Hz, 2H), 2.86 (t, J=7.1 Hz, 2H), 3.03 (t, J=7.6 Hz, 2H); ¹³C NMR(125 MHz, D₂O) δ 19.1, 21.5, 22.0, 24.6, 34.1, 39.1, 48.2, 60.2; ES-MS236 (M+H)

3-[(1-methylcycloheptyl)amino]-1-propanesulfonic acid (Compound FS)

For the Ritter reaction, the flask was closed with a septum andconnected to a 20% NaOH scrubber. Potassium cyanide (powdered, 2.8 g, 43mmol) was added in portions to acetic acid (10 mL). The mixture wasstirred for 10 minutes at room temperature. A solution of sulfuric acid(7 mL) in acetic acid (7 mL) was added drop-wise over a 20 minuteperiod. Then, the 1-methyl-1-cycloheptanol (5 g, 39.0 mmol) was addeddrop-wise over 5 minutes. The mixture was stirred at room temperaturefor 22 h then cooled to 0° C. with a ice/water bath. The pH of thesolution was adjusted to 9 with the addition of 50% NaOH (about 70 g).The layers were separated and the aqueous layer was extracted with ether(1×20 mL). The combined organic layers were washed with saturated sodiumcarbonate (1×5 mL) then dried over sodium sulfate. The ether wasevaporated under reduced pressure to afford a clear yellow oil (5.71 g,94%). The oil showed to be a mixture of cis and trans formamide but whatotherwise pure enough to be used as such. ¹H NMR (500 MHz, CDCl₃) δ 1.34(s, 1.5H), 1.43 (s, 1.5H), 1.49-1.60 (m, 8H), 1.96-2.00 (m, 1H), [5.28and 5.95 (br s, 1H)], [8.06 (s) and 8.28 (d, J=12.2 Hz) for 1H]; ¹³C NMR(125 MHz, CDCl₃) δ 22.2, 22.4, 27.7, 29.3, 29.4, 30.7, 40.5, 42.5, 56.0,57.4, 160.5, 163.3

A solution of NaOH (25%, 40 mL) was added to the crude1-methyl-1-cycloheptylformamide (5.7 g). The mixture was heated toreflux for 3 hours then cooled at room temperature. Some sodium chloride(7.5 g) was added to facilitate the phase separation. The layers wereseparated and the aqueous layer was extracted with MTBK (1×10 mL). Thecombined organic layers were washed with brine (1×5 mL) the dried oversodium sulfate and filtered. The filtrate was used as such in the nextstep. ¹H NMR (500 MHz, CD₃OD) δ 1.10 (s, 3H), 1.40-1.48 (m, 2H),1.5-1.65 (m, 10H); ¹³C NMR (125 MHz, CD₃OD) δ 24.0, 31.2, 31.4, 44.4,53.6.

A solution of 1,3-propanesultone (4.3 g, 35 mmol) in toluene (10 mL) wasadded dropwise to a the crude solution of 1-methyl-1-cycloheptylamine inMTBK (total volume 30 mL). The mixture was heated to reflux for 18 hoursthen was cooled to room temperature. The solid was collected by suctionfiltration, rinsed with acetone (2×5 mL). The solid was dried overnightat 45° C. in the vacuum oven. The title compound was obtained as a finewhite solid (7.77 g, 31.2 mmol, 80% overall yield). ¹H NMR (500 MHz,D₂O) δ 1.27 (s, 3H), 1.40-1.60 (m, 8H), 1.71-1.81 (m, 4H), 2.00-2.06 (m,2H), 2.95 (t, J=6.3 Hz, 2H), 3.13 (t, J=7.1 Hz, 2H); ¹³C NMR (125 MHz,D₂O) δ 22.0, 22.1, 23.3, 29.5, 37.1, 40.0, 48.3, 64.0; ES-MS 250 (M+H)

Preparation of3-{[(1R)-1-(benzyloxycarbonyl)-3-methylbutyl]amino}-1-propanesulfonicacid (Compound HI)

D-Leucine benzylester p-tosylate (2.5 g, 6.3 mmol) was treated with anaqueous solution of K₂CO₃ (30 mL). The mixture was extracted with EtOAc(3×30 mL). The organic extracts were separated, combined, dried withNa₂SO₄, filtered, evaporated under reduced pressure and dried in vacuo.

To a solution of D-Leucine benzylester (6.3 mmol) in acetonitrile (9 mL)and MeOH (3 mL) was added 1,3-propane sultone (691 mg, 5.7 mmol). Thesolution was stirred at reflux for 2.5 hours. The reaction mixture wascooled to room temperature. The solid material was filtered and washedwith aconitrile (2×20 mL). The solid was dissolved in 20% water/EtOH (75mL). Dowex Marathon C ion exchange resin (strongly acidic) was added tothe solution. The suspension was stirred for 15 minutes before the resinwas removed by filtration. The filtrate was evaporated under reducedpressure and dried in vacuo, affording the title compound (960 mg, 49%).¹H NMR (D₂O, 500 MHz) δ ppm 7.52 (m, 5H), 5.41 (d, 1H, J=12.2 Hz), 5.35(d, 1H, J=12.2 Hz), 4.16 (m, 1H), 3.22 (m, 2H), 2.97 (t, 2H, J=6.8 Hz),2.16 (m, 2H), 1.88 (m, 1H), 1.79 (m, 1H), 1.76 (m, 1H), 0:94 (d, 6H,J=3.9 Hz). ¹³C (DMSO, 125 MHz) δ ppm 169.60, 135.62, 129.24, 129.21,129.11, 68.08, 58.09, 49.87, 46.48, 24.77, 23.50, 22.50, 22.04.[α]_(D)=−2.1° (c=0.00095 in water), ES-MS 344 (M+1).

Preparation of 3-[(5-hydroxy-1,5-dimethylhexyl)amino]-1-propanesulfonicacid (Compound HJ)

To a solution of 6-amino-2-methyl-2-heptanol (2.5 g, 17.2 mmol) inacetonitrile (22 mL) was added 1,3-propane sultone (2.0 g, 16.4 mmol).The solution was stirred at reflux for 2 hours. The reaction mixture wascooled to room temperature. The solid material was filtered and washedwith acetonitrile (2×20 mL). The solid was dissolved in 20% MeOH (75mL). Dowex Marathon C ion exchange resin (strongly acidic) was added tothe solution. The suspension was stirred for 15 minutes before the resinwas removed by filtration. The filtrate was evaporated under reducedpressure. The solid was suspended in acetone (150 mL), and then thesolid material was filtered and dried in vacuo, affording the titlecompound (3.08 g, 70%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.19 (m, 1H), 3.08(m, 2H), 2.88 (t, 2H, J=7.3 Hz), 1.99 (m, 2H), 1.60 (m, 2H), 1.36 (m,4H), 1.18 (d, 3H, J=6.8 Hz), 1.07 (s, 6H). ¹³C (D₂O, 125 MHz) δ ppm71.63, 54.73, 48.08, 43.46, 42.27, 32.97, 27.78, 27.73, 21.64, 19.67,15.43. ES-MS 268 (M+1).

Preparation of3-{[(1R)-2-methoxy-1-methyl-2-oxoethyl]amino}-1-propanesulfonic acid(Compound HK)

D-Alanine methylester hydrochloride (3.0 g, 21.5 mmol) was treated witha aqueous solution of K₂CO₃ (50 mL). The mixture was extracted withEtOAc (3×50 mL). The organic extracts were separated, combined, driedwith Na₂SO₄, filtered and evaporated under reduced pressure.

To a solution of D-Alanine methylester (1.33 g, 12.9 mmol) inacetonitrile (15 mL was added 1,3-propane sultone (1.42 g, 11.7 mmol).The solution was stirred at reflux for 2 hours. The reaction mixture wascooled to room temperature. The solid material was filtered and washedwith acetonitrile (2×15 mL). The solid was dissolved in water (30 mL).Dowex Marathon C ion exchange resin (strongly acidic) was added to thesolution. The suspension was stirred for 15 minutes before the resin wasremoved by filtration. The filtrate was evaporated under reducedpressure and dried in vacuo, affording the title compound (1.52 g, 42%).¹H NMR (D₂O, 500 MHz) δ ppm 4.07 (m, 1H), 3.72 (s, 3H), 3.14 (m, 2H),2.89 (t, 2H, J=7.3 Hz), 2.03 (m, 2H), 1.46 (dd, 3H, J=1.95 Hz, 7.3 Hz).¹³C (DMSO, 125 MHz) δ ppm 170.74, 55.62, 53.82, 47.96, 44.76, 21.53,14.03. [α]_(D)=+1.4° (c=0.0088 in water), ES-MS 224 (M−1).

Preparation of 4-(1,2,3,4-tetrahydro-1-naphthylamino)-2-butanesulfonicacid (Compound JF)

To a solution of 1,2,3,4-tetrahydro-1-naphthylamine (2.01 g, 13.6 mmol)in 2-butanone (15 mL) was added 2,4-butane sultone (L⁹⁵ g, 14.3 mmol).The solution was stirred at for 2 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration, washed with acetone(2×25 mL) and dried in vacuo. ¹H NMR (DMSO, 500 MHz) δ ppm 8.56 (s(broad), 1H), 7.49 (dd, 1H, J=8.0 Hz, 11.9 Hz), 7.29 (m, 1H), 7.26 (m,1H), 7.19 (d, 1H, J=8.0 Hz), 4.40 (d, 1H, J=13.7 Hz), 3.14 (m, 2H), 2.75(m, 3H), 1.96 (m, 5H), 1.40 (m, 1H), 1.23 (m, 3H). ¹³C (DMSO, 125 MHz) δppm 138.81, 131.81, 130.40, 130.28, 130.16, 129.41, 126.76, 126.73,54.97, 54.58, 54.08, 44.18, 43.23, 29.50, 28.84, 24.84, 24.76, 18.23,18.20, 17.79, 17.01. ES-MS 284 (M+1).

Preparation of 4-(octylamino)-2-butanesulfonic acid (Compound JG)

To a solution of octylamine (2.00 g, 15.5 mmol) in 2-butanone (17 mL)was added 2,4-butane sultone (2.21 g, 16.2 mmol). The solution wasstirred at reflux for 2 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration, washed with acetone(2×25 mL) and dried in vacuo. It was suspended in a solution of 25%EtOH/acetone (50 mL). The suspension was stirred for 5 minutes. Thesolid was collected by filtration, washed with acetone (2×25 mL) anddried in vacuo. ¹H NMR (DMSO, 500 MHz) δ ppm 8.45 (s (broad), 1H), 3.01(m, 1H), 2.84 (m, 2H), 2.58 (m, 1H), 1.92 (m, 1H), 1.75 (m, 1H), 1.51(m, 2H), 1.10 (d, 1H, J=6.8 Hz), 0.85 (t, 3H, J=6.8 Hz). ¹³C (DMSO, 125MHz) δ ppm 53.05, 47.27, 46.15, 31.83, 29.42, 29.13, 26.51, 26.27,22.75, 17.19, 14.64. ES-MS 266 (M+1).

Preparation of 4-(adamantyl)amino-2-butanesulfonic acid (Compound JH)

1-adamantaneamine hydrochloride (2.51 g, 13.3 mmol) was treated with 1NNaOH (20 mL) and CH₂Cl₂ (3×20 mL). The organic extracts were combined,dried with Na₂SO₄, filtered, evaporated under reduced pressure and driedin vacuo.

To a solution of 2-adamantanamine (1.99 g, 13.1 mmol) in acetonitrile(15 mL) was added 2,4-butane sultone (1.87 g, 13.8 mmol). The solutionwas stirred at reflux for 2 hours.

The reaction was cooled to room temperature. The solid was collected byfiltration, washed with acetonitrile (3×25 mL) and dried in vacuo. ¹HNMR (DMSO, 500 MHz) δ ppm 8.53 (s (broad), 1H), 3.33 (m, 2H), 2.61 (m,1H), 2.10 (s, 3H), 1.93 (m, 1H), 1.77 (m, 7H), 1.61 (m, 6H), 1.12 (d,1H, J=6.8 Hz). ¹³C (DMSO, 125 MHz) δ ppm 56.20, 53.34, 35.85, 29.75,29.04, 17.206. ES-MS 288 (M+1).

Preparation of 4-(2-adamantyl)amino-2-butanesulfonic acid (Compound JI)

The 2-adamantanamine hydrochloride (2.50 g, 13.3 mmol) was treated with1N NaOH (20 mL) and CH₂Cl₂ (3×20 mL). The organic extracts werecombined, dried with Na₂SO₄, filtered, evaporated under reduced pressureand dried in vacuo.

To a solution of 1-adamantanamine (1.99 g, 13.1 mmol) in acetonitrile(15 mL) was added 2,4-butane sultone (L⁸⁷ g, 13.8 mmol). The solutionwas stirred at reflux for 2 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration, washed withacetonitrile (2×25 mL) and dried in vacuo. ¹H NMR (DMSO, 500 MHz) δ ppm3.20 (m, 1H), 3.05 (m, 2H), 2.67 (m, 1H), 2.07 (m, 2H), 2.00 (m, 1H),1.95 (m, 4H), 1.82 (m, 4H), 1.69 (m, 4H), 1.55 (m, 4H), 1.12 (d, 1H,J=8.0 Hz). ¹³C (DMSO, 125 MHz) δ ppm 62.27, 53.91, 44.33, 37.30, 36.82,36.77, 30.30, 30.20, 29.57, 29.50, 28.95, 17.12, 26.85, 17.44. ES-MS 288(M+1).

Preparation of 4-(bicyclo[2.2.1]hept-2-ylamino)-2-butanesulfonic acid(Compound JJ)

To a solution of exo-2-aminonorbornane (1.0 g, 9.0 mmol) intetrahydrofuran (THF, 10 mL) was added 2,4-butane sultone (1.28 g, 9.3mmol). The solution was stirred at reflux for 3 hours. The reaction wascooled to room temperature. The solid was collected by filtration,washed with THF (2×20 mL) and dried in vacuo. ¹H NMR (DMSO, 500 MHz) δppm 8.43 (s (broad), 1H), 2.96 (m, 3H), 2.62 (m, 1H), 2.38 (m, 1H),2.28, (m, 1H), 1.91 (m, 1H), 1.82 (m, 1H), 1.61 (m, 1H), 1.54 (m, 2H),1.42 (m, 2H), 1.12 (m, 6H). ¹³C (DMSO, 125 MHz) δ ppm 60.92, 60.79,53.61, 53.21, 44.55, 44.36, 39.80, 39.55, 36.27, 36.15, 36.11, 35.98,35.19, 35.13, 29.62, 29.43, 28.07, 26.88, 17.56, 14.11. ES-MS 248 (M+1).

Preparation of 4-(azoniabicyclo[2.2.2]oct-2-ylamino)-2-butanesulfonate(Compound JK)

Quinuclidine hydrochloride (2.50 g, 16.9 mmol) was treated with 1N NaOH(20 mL) and CH₂Cl₂ (4×20 mL). The organic extracts were combined, driedwith Na₂SO₄, filtered, evaporated under reduced pressure and dried invacuo.

To a solution of quinuclidine (900 g, 8.2 mmol) in tetrahydrofuran (THF,18 mL) and MeOH (0.5 mL) was added 2,4-butane sultone (1.16 g, 8.6mmol). The solution was stirred at reflux overnight. The reaction wascooled to room temperature. The solid was collected by filtration,washed with THF (2×25 mL) and dried in vacuo. ¹H NMR (DMSO, 500 MHz) δppm 3.40 (m, 7H), 3.20 (td, 1H, J=3.9 Hz, 12.7 Hz), 2.38 (m, 1H), 2.01(m, 2H), 1.83, (m, 6H), 1.70 (m, 1H), 1.10 (d, 3H, J=12.7 Hz). ¹³C(DMSO, 125 MHz) δ ppm 62.63, 54.23, 52.18, 25.67, 24.07, 19.78, 17.04.ES-MS 208 (M+1).

Preparation of 4-[(dl)-1-hydroxy-2-pentyl]amino-2-butane sulfonic acid(Compound JL)

To a solution of DL-2-aminopentanol (1.0 g, 9.7 mmol) in tetrahydrofuran(11 mL) was added 2,4-butane sultone (1.45 g, 10.2 mmol). The solutionwas stirred at reflux for 4 hours. The reaction was cooled to roomtemperature. The supernatant was removed and the solid was dried invacuo. The product was suspended in 2-propanol (100 mL) and the mixturewas stirred for 5 minutes. The white solid was filtered, washed with2-propanol and dried in vacuo. ¹H NMR (D₂O, 500 MHz) δ ppm 3.74 (d, 1H,J=12.7 Hz), 3.59 (dd, 1H, J=5.4 Hz, 12.9 Hz), 3.13 (m, 3H), 2.89 (m,1H), 2.07 (m, 1H), 1.82, (m, 1H), 1.52 (m, 2H), 1.28 (m, 2H), 1.18 (d,3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 59.43, 58.80, 53.47, 42.88,29.30, 28.43, 18.40, 14.99, 13.23. ES-MS 240 (M+1).

Preparation of 4-(nonylamino)-2-butanesulfonic acid (Compound JN)

To a solution of nonylamine (2.00 g, 14.0 mmol) in tetrahydrofuran (THF,15 mL) was added 2,4-butane sultone (2.08 g, 14.7 mmol). The solutionstirred at reflux for 5 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration, washed with THF(2×25 mL) and dried in vacuo.

The product (1.10 g, 3.9 mmol) was dissolved with heating in a solutionof EtOH (20 mL), water (600 uL) and NaOH (163 mg, 4.1 mmol). After a fewminutes a white solid precipitated. The solid was collected byfiltration, washed with acetone (2×25 mL) and dried in vacuo. ¹H NMR(D₂O, 500 MHz) δ ppm 2.70 (m, 1H), 2.52 (m, 2H), 2.37 (m, 1H), 1.95 (m,1H), 1.46 (m, 1H), 1.34 (m, 1H), 1.14 (m, 17H), 0.70 (t, 3H, J=6.8 Hz).¹³C (D₂O, 125 MHz) δ ppm 54.31, 52.90, 50.44, 31.31, 28.80, 28.74,28.57, 27.31, 26.95, 22.20, 14.60, 13.57. ES-MS 302 (M+1).

Preparation of 4-(dimethylamino)-2-butanesulfonic acid (Compound JO)

2,4-butanesultone (1.27 g, 8.9 mmol) was added to an ice-chilledsolution of dimethylamine (40% w/w in water). The solution was stirredat 0° C. for 4 hours. The solvent was evaporated in vacuo until completedryness. The solid was washed with acetone (50 mL), collected byfiltration and dried in vacuo. ¹H NMR (D₂O, 500 MHz) δ ppm 3.18 (t, 2H,J=8.1 Hz), 2.85 (m, 1H), 2.76 (s, 6H), 2.09 (m, 1H), 1.81 (m, 1H), 1.17(d, 3H, J=7.3 Hz). ¹³C (D₂O, 125 MHz) δ ppm 55.65, 53.05, 42.88, 26.72,14.81. ES-MS 182 a (M+1).

Preparation of 4-(benzylamino)-2-butanesulfonic acid, sodium salt(Compound JP)

To a solution of benzylamine (1.50 g, 14.0 mmol) in tetrahydrofuran(THF, 18 mL) was added 2,4-butane sultone (1.98 g, 14.6 mmol). Thesolution was stirred at reflux for 2 hours. The reaction was cooled toroom temperature. The solid was collected by filtration, washed with THF(2×25 mL) and dried in vacuo.

The product (2.55 g, 10.5 mmol) was dissolved with heating in a solutionof EtOH (25 mL), water (1.6 mL) and NaOH (440 mg, 11.0 mmol). Diethylether (150 mL) was added to the filtrate. The solid was filtered anddried in vacuo. Yield: 27%. ¹H NMR (DMSO, 500 MHz) δ ppm 7.29 (m, 4H),7.20 (m, 1H), 3.67 (m, 2H), 2.56 (m, 1H), 2.45 (m, 2H), 1.98 (m, 1H),1.36 (m, 1H), 1.04 (d, 3H, J=6.8 Hz). ¹³C (DMSO, 125 MHz) δ ppm 141.49,128.73, 128.61, 127.16, 53.49, 52.88, 47.31, 32.67, 16.53. ES-MS 266(M+1).

Preparation of 4-(ethylamino)-2-butanesulfonic acid, sodium salt(Compound JQ)

A solution of 2,4-butanesultone (1.33 g, 9.3 mmol) in tetrahydrofuran(THF, 3.0 mL) was added via syringe pump over a 2 h period to ethylamine(70% w/w in water, 12.0 mL, 186.0 mmol) at 5° C. The solution wasstirred at 5° C. for an additional 2 hours. The solvent wasco-evaporated with EtOH (3×25 mL). The solid was suspended in acetone(25 mL). The suspension was stirred for 5 minutes the solid wasfiltered, washed with acetone (2×25 mL) and dried in vacuo. Yield: 70%.¹H NMR (D₂O, 500 MHz) δ ppm 3.06 (t, 2H, J=8.1 Hz), 2.97 (m, 2H), 2.87(m, 2H), 2.06 (m, 1H), 1.77 (m, 1H), 1.18 (d, 3H, J=7.3 Hz), 1.14 (t,3H, J=7.3 Hz). ¹³C (D₂O, 125 MHz) δ ppm 53.16, 44.91, 43.03, 28.20,14.76, 10.66. ES-MS 182 (M+1).

Preparation of 4-(tert-butylamino)-1-butanesulfonic acid (Compound LD)

To a solution of tert-butylamine (1.0 mL, 9.5 mmol) in tetrahydrofuran(4 mL) was added 1,4-butane sultone (1.36 g, 10.0 mmol) at roomtemperature. The solution was stirred at reflux for 2 hours. Thereaction was cooled to room temperature. The solid was collected byfiltration, washed with acetone (2×20 mL) and dried in vacuo. Yield: 690mg (34%). ¹H NMR (D₂O, 500 MHz) δ ppm 2.92 (t, 2H, J=7.1 Hz), 2.82 (t,2H, J=7.1 Hz), 1.68 (m, 4H), 1.22 (s, 9H). ¹³C (D₂O, 125 MHz) δ ppm57.07, 50.30, 40.95, 25.28, 24.96, 21.62. ES-MS 210 (M−1).

Preparation of 4-amino-2-butanesulfonic acid (Compound JR)

A solution of 2,4-butanesultone (1.0 g, 7 mmol) in tetrahydrofuran (THF,4.0 mL) was added via syringe pump over a 4 h period to ammoniumhydroxide (28-30% NH₃, 43 mL, 350 mmol) at 5° C. The solution wasstirred at 5° C. for an additional 30 minutes. The solvent wasco-evaporated with EtOH (3×25 mL). The solid was dried in vacuo. Yield:94%. ¹H NMR (D₂O, 500 MHz) δ ppm 3.05 (m, 2H), 2.90 (m, 1H), 2.05 (m,1H), 2.06 (m, 1H), 1.77 (m, 1H), 1.18 (d, 3H, J=6.8 Hz), 1.14 (t, 3H,J=7.3 Hz). ¹³C (DMSO, 125 MHz) δ ppm 52.75, 38.20, 30.87, 17.27. ES-MS154 (M+1).

Preparation of 4-piperidin-1-yl-2-butanesulfonic acid (Compound JS)

To a solution of piperidine (1.50 g, 17.6 mmol) in tetrahydrofuran (THF,20 mL) was added 2,4-butanesultone (2.50 g, 18.5 mmol). The solution wasstirred at reflux for 3 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration, washed with THF(2×20 mL) and dried in vacuo.

The product (3.53 g, 15.9 mmol) was dissolved with heating in a solutionof EtOH (30 mL), water (1.3 mL) and NaOH (670 mg, 16.7 mmol). Thesolution was poured in a large excess of Et₂O (500 mL). The solid wasfiltered, washed with Et₂O (1×25 mL) and acetone (1×20 mL) and dried invacuo. Yield: 64%. ¹H NMR (D₂O, 500 MHz) δ ppm 2.72 (m, 1H), 2.33 (m,6H), 1.97 (m, 1H), 1.48 (m, 1H), 1.43 (m, 4H), 1.31 (m, 1H), 1.13 (d,3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 55.78, 54.42, 53.61, 27.73,24.92, 23.59, 14.61. ES-MS 244 (M+1).

Preparation of 4-(ethylamino)-1-butanesulfonic acid (Compound LE)

A solution of 1,4-butanesultone (2.66 g, 18.6 mmol) in tetrahydrofuran(total volume: 4 mL) was added via syringe pump over a 4 hour period toethylamine (70% w/w in water, 24 mL, 372 mmol) at 5° C. The solution wasstirred at 5° C. for an additional 3 hours before it was warm up to roomtemperature. The reaction was stirred in these conditions overnight. Thesolvent was co-evaporated with EtOH (1×25 mL). The solid was suspendedin 50% acetone/EtOH (50 mL). The suspension was stirred for 5 minutes,the solid was filtered and dried in vacuo. Yield: 75%. ¹H NMR (D₂O, 500MHz) δ ppm 2.95 (m, 4H), 2.82 (m, 2H), 1.68 (m, 4H), 1.13 (t, 3H, J=7.3Hz), 1.14 (t, 3H, J=7.3 Hz). ¹³C (D₂O, 125 MHz) δ ppm 50.27, 46.68,42.99, 24.66, 21.48, 10.64. ES-MS 182 (M+1).

Preparation of 4-(azoniabicyclo[2.2.2]oct-2-ylamino)-1-butanesulfonate(Compound LF)

To a solution of quinuclidine (1.5 g, 13.5 mmol) in tetrahydrofuran(THF, 15 mL) was added 1,4-butanesultone (2.0 g, 14.4 mmol) at roomtemperature. The solution was stirred at reflux for 2 hours. Thereaction was cooled to room temperature. The solid was collected byfiltration, washed with THF (2×25 mL) and dried in vacuo. ¹H NMR (D₂O,500 MHz) δ ppm 3.26 (m, 6H), 3.02 (m, 2H), 2.82 (t, 2H, J=17.3 Hz), 2.04(m, 1H), 1.84, (m, 6H), 1.75 (m, 2H), 1.64 (m, 2H). ¹³C (D₂O, 125 MHz) δppm 63.68, 54.81, 50.14, 23.51, 21.45, 20.58, 19.19. ES-MS 248 (M+1).

Preparation of 3-(dimethylamino)-2-hydroxy-1-propane sulfonic acid,sodium salt (Compound JT)

A solution of 3-chloro-2-hydroxy-1-propanesulfonic acid, sodium salt (10g, 48.3 mmol) in water (40 mL total volume) was added via a syringe pumpover 4 hours to a cold (2.8-3.1° C.) solution of dimethylamine (40% wtin water, 300 mL) with stirring. The mixture was slowly warmed to roomtemperature overnight. The mixture was then co-evaporated with absoluteethanol (20 mL) and concentrated to dryness. The solid was driedovernight at 60° C. in the vacuum oven. The solid was suspended inethanol (40 mL) stirred at reflux for 2 hours. The suspension was cooledto 5° C. and the solid was collected by suction-filtration,aspirator-dried 5 minutes, then dried for the weekend at 60° C. in thevacuum oven (wet cake: 13.74 g). The desired material was obtained as anoff-white solid (11.65 g, quantitative).

Preparation of 4-Dimethylamino-1-butanesulfonic acid (Compound LH)

A solution of 1,4-butane sultone (7.5 mL, 73.6 mmol) in 1,4-dioxane(total volume: 10 mL) was added over 4 hours via a syringe pump to acold (4.3° C.) solution of dimethylamine (40% wt in water, 275 mL). Themixture was stirred for 3 hours at 4° C. after the end of the addition,then overnight at room temperature. The mixture was concentrated todryness. The solid was suspended in absolute ethanol (50 mL) and themixture was heated to reflux for 90 minutes. The suspension was cooledto 5° C. and the solid was collected by suction-filtration, rinsed withethanol (2×10 mL). The solid was dried for 18 h at 60° C. in the vacuumoven. The desired material was obtained as a fine white powder 13.21 g,72.9 mmol, 99% yield. The ¹H and ¹³C NMR and MS were consistent with thestructure.

Preparation of 3-(ethylamino)-2-hydroxy-1-propanesulfonic acid (CompoundJU)

A solution of 3-chloro-2-hydroxy-1-propanesulfonic acid sodium salt (10g, 50.9 mmol) in water (total volume: 40 mL) was added over 5 hours, viaa syringe pump, to a cold (4.7° C.) solution of ethyl amine in water.The mixture was stirred for an additional 2 hours at 4.7° C. then for 18hours at room temperature. NMR: quantitative yield. The mixture wasconcentrated. A solid could not be obtained: the sodium salt was toohygroscopic. The solution was treated with Amberlite IR-120 Plus, acidform, ion-exchange resin to give the free acid. It was still toohygroscopic to be obtained as a solid form. Submitted as a solution:d=1.314 g/mL, 62.5% w/w of the free acid in water. The ¹H and ¹³C NMRand MS were consistent with the structure.

Preparation of 3-(tert-butylamino)-2-hydroxy-1-propanesulfonic acid(Compound JV)

A solution of 3-chloro-2-hydroxy-1-propanesulfonic acid, sodium salt (15g, 25 mmol) in water (12 mL total volume) was added over 5 minutes to amixture of tert-butylamine (12.5 mL), water (6 mL) and methanol (3 mL).The mixture was heated at 35° C. for 1 h, 40° C. for 1 h, 45° C. for 1.5hours. The mixture was the concentrated to a thick oil. The crudereaction mixture was passed over a column of Dowex 50×8 (125 g). Thefractions containing the product were concentrated to dryness. The solidwas dried overnight at 60° C. in the vacuum oven. The solid wasrecrystallized in a mixture of methanol (25 mL) and water (7 mL). Themixture was cooled slowly to room temperature, then to 5° C. The solidwas collected by suction-filtration, rinsed with ethanol (1×10 mL). Thesolid was then dried for 18 hours at 60° C. in the vacuum oven. Thedesired material was obtained as an off-white solid (3.11 g, 14.7 mmol,59%). The ¹H and ¹³C NMR and MS were consistent with the structure.

Preparation of 1-(N-octylamino)-2-hydroxy-1-propanesulfonic acid(Compound JW)

A solution of 3-chloro-2-hydroxy-1-propanesulfonic acid, sodium salt (4g, 20 mmol) in water (17.5 mL total volume) was added over 2 hours to amixture of octylamine (8 mL), water (20 mL) and 1,4-dioxane (11 mL) at70-75° C. The mixture was stirred at this temperature for another 2hours after the end of the addition. The 1,4-dioxane was removed underreduced pressure and the mixture was diluted with water (10 mL). Themixture was extracted with 40% ethyl acetate/hexane (3×40 mL). Theaqueous layer was concentrated then the mixture was passed over a columnof Dowex 50×8 (125 g). The fractions containing the pure product wereconcentrated to a thick oil then freeze-dried. The desired material wasobtained as a white fluffy solid (150 mg, 0.56 mmol, 3%). The ¹H and ¹³CNMR and MS were consistent with the structure.

Preparation of 1-(3-sulfo-2-hydroxypropyl) quinuclidinium, inner salt(Compound JX)

A solution of 3-chloro-2-hydroxy-1-propanesulfonic acid, sodium salt (2g, 10 mmol) in water (12 mL total volume) was added over 1 hour to amixture of quinuclidine (1.63 g, 4.7 mmol), water (10 mL) and1,4-dioxane (10 mL) at 80° C. The mixture was stirred at thistemperature for another 2 hours after the end of the addition. Thereaction mixture was concentrated then the mixture was passed over acolumn of Dowex 50×8 (125 g). The fractions containing the pure productwere concentrated to a white solid. The solid was dried for 18 hours at60° C. in the vacuum oven. The desired material was obtained as a whitesolid (1.92 g, 7.7 mmol, 77%). The ¹H and ¹³C NMR and MS were consistentwith the structure.

Preparation of 1-(N-benzylamino)-2-hydroxy-1-propanesulfonic acid,sodium salt (Compound JY)

A solution of 3-chloro-2-hydroxy-1-propanesulfonic acid, sodium salt (4g, 20 mmol) in water (12.5 mL total volume) was added over 2 hours to amixture of benzylamine (4.28 g, 40 mmol), water (10 mL) and 1,4-dioxane(5 mL) at 80° C. The mixture was stirred at this temperature for another2.5 hours after the end of the addition. The reaction mixture wasextracted with chloroform (2×40 mL). It was then concentrated todryness. The crude solid was recrystallized in a mixture of ethanol (30mL) and water (4 mL). The mixture was left to cool to room temperaturefor the night. The solid was collected by suction-filtration, rinsedwith ethanol (10 mL) and dried in the vacuum oven at 60° C. The desiredmaterial was obtained as a white solid (2.67 g, 10 mmol, 50%). The ¹Hand ¹³C NMR and MS were consistent with the structure.

Preparation of2-hydroxy-3-(1,2,3,4-tetrahydronaphthalen-1-ylamino)propane-1-sulfonicacid (Compound JZ)

A solution of 3-chloro-2-hydroxy-1-propanesulfonic acid, sodium salt (2g, 10 mmol) in water (9.75 mL total volume) was added over 8 hours to amixture of 1,2,3,4-tetrahydro-1-nahtylamine (2 g, 13.6 mmol), water (10mL) and 1,4-dioxane (4 mL) at 40° C. The mixture was stirred at thistemperature for another 18 hours after the end of the addition. Thereaction was not completed. The mixture was heated for 2 hours atreflux. The mixture was diluted with water (10 mL) and 50% w/w NaOH(0.25 mL) was added. The reaction mixture was extracted with chloroform(2×25 mL). It was then concentrated to a thick oil. The solution wasapplied on a Dowex 50 W 8 column (100 g). The fractions containing theproduct were concentrated, treated with activated charcoal (no effect)and freeze-dried. The desired material was obtained as a glassy solid(0.85 g, 3 mmol, 30%). The ¹H and ¹³C NMR and MS were consistent withthe structure.

Preparation of 2-hydroxy-3-piperidin-1-ylpropane-1-sulfonic acid(Compound KA)

A solution of 3-chloro-2-hydroxy-1-propanesulfonic acid, sodium salt (4g, 20 mmol) in water (13.35 mL total volume) was added over 5 hours to asolution of piperidine (8 mL g, 80 mmol), in water (15 mL) at 70° C. Themixture was stirred at 80° C. for 2 hours. The reaction was completed.The mixture was stirred at room temperature for the night. The mixturewas diluted with water (10 mL) and was extracted with chloroform (3×30mL). It was then concentrated to a thick oil. The solution was appliedon a Dowex 50 W 8 column (100 g). The fractions containing the productwere concentrated to dryness then recrystallized in a mixture of ethanol(30 mL) and water (2.1 mL). The mixture was cooled slowly at roomtemperature. The solid was collected by suction filtration, rinsed withethanol (2×5 mL) air dried 5 minutes, then 18 hours at 60° C. in thevacuum oven. The desired material was obtained as a fine white solid(3.06 g, 13.7 mmol, 68%). The ¹H and ¹³C NMR and MS were consistent withthe structure.

Preparation of 4-(adamantyl)amino-1-butanesulfonic acid (Compound LI)

1-adamantaneamine hydrochloride (2.67 g, 13.3 mmol) was treated with 1NNaOH (20 mL) and CH₂Cl₂ (3×20 mL). The biphasic solution was shaken. Theorganic extracts were combined, dried with Na₂SO₄, filtered, evaporatedunder reduced pressure and dried in vacuo.

To a solution of 2-adamantanamine (1.87 g, 12.4 mmol) in tetrahydrofuran(THF, 15 mL) was added 1,4-butane sultone (1.76 g, 13.0 mmol). Thesolution was stirred at reflux overnight. The reaction was cooled toroom temperature. The solid was collected by filtration, washed with THF(1×15 mL) and dried in vacuo. A suspension of the solid in EtOH (25 mL)was stirred at reflux for 1 hour. The warm mixture was filtered. Thesolid was dried in vacuo. ¹H NMR (D₂O, 500 MHz) δ ppm 2.92 (m, 2H), 2.82(m, 1H), 2.05 (s, 3H), 1.75 (s, 6H), 1.63 (m, 6H), 1.52 (m, 3H). ¹³C(D₂O, 125 MHz) δ ppm 57.62, 50.30, 39.03, 38.14, 35.09, 28.98, 25.25,21.63. ES-MS 288 (M+1).

Preparation of 4-(octylamino)-1-butanesulfonic acid (Compound LJ)

To a solution of octylamine (2.20 g, 17.0 mmol) in tetrahydrofuran (11mL) was added 1,4-butane sultone (2.30 g, 16.2 mmol). The solution washeated to reflux for 5 hours. The reaction was cooled to roomtemperature. The product formed a gel. A few drops of EtOH were added todissolve the product. The solution was poured in a large excess ofacetone (25 mL). After 5 minutes, a white solid precipitated. The solidwas collected by filtration and dried in vacuo. The product wasdissolved in EtOH and Dowex 50×8 resin (pre-washed, 6 g) was added tothe solution. The suspension was stirred for 15 minutes and the resinwas filtered. The filtrate was evaporated under educed pressure and theproduct was dried in vacuo. Yield: 31%. ¹H NMR (DMSO, 500 MHz) δ ppm8.24 (s (broad), 1H), 2.85 (m, 4H), 2.45 (m, 2H), 1.64 (m, 4H), 1.61 (m,2H), 1.25 (m, 10H), (m, 2H), 0.85 (t, 3H, J=6.8 Hz). ¹³C (DMSO, 125 MHz)δ ppm 51.22, 47.54, 47.38, 31.82, 29.14, 26.59, 26.13, 25.51, 22.95,22.75, 14.64. ES-MS 266 (M+1).

Preparation of 4-(cyclohexylamino)-2-butanesulfonic acid (Compound KM)

To a solution of cyclohexylamine (1.50 g, 15.1 mmol) in tetrahydrofuran(15 mL) was added 2,4-butane sultone (2.04 g, 14.4 mmol). The solutionstirred at reflux for 2 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration and dried in vacuo.Yield: 59%. ¹H NMR (DMSO, 500 MHz) δ ppm 8.50 (s (broad), 1H), 3.02 (m,2H), 2.93 (m, 1H), 2.60 (m, 1H), 1.93 (m, 3H), 1.75 (m, 3H), 1.57 (m,1H), 1.21 (m, 4H), 1.11 (m, 4H). ¹³C (DMSO, 125 MHz) δ ppm 56.23, 53.20,43.09, 29.54, 29.41, 29.39, 25.40, 24.42, 17.23. ES-MS 234 (M−1).

Preparation of 4-[(dl)-1-hydroxy-2-pentyl]amino-1-butanesulfonic acid(Compound LL)

To a solution of DL-2-aminopentanol (1.0 g, 9.7 mmol) in tetrahydrofuran(6 mL) was added 1,4-butane sultone (1.31 g, 9.2 mmol) at roomtemperature. The solution was stirred at reflux for 5 hours. Thereaction was cooled to room temperature. The supernatant was removed andthe solid was dried in vacuo. The white solid was filtered, washed withacetone (2×25 mL) and dried in vacuo. Yield: 45%. ¹H NMR (DMSO, 500 MHz)δ ppm 8.20 (s (broad), 1H), 5.23 (m, 1H), 3.66 (m, 1H), 3.49 (m, 1H),3.02 (m, 1H), 2.91 (m, 2H), 2.46 (t, 2H, J=7.3 Hz), 1.65, (m, 4H), 1.54(m, 2H), 1.38 (m, 2H), 0.88 (t, 3H, J=7.3 Hz). ¹³C (DMSO, 125 MHz) δ ppm58.83, 58.54, 51.23, 44.77, 29.91, 25.56, 23.06, 18.95, 14.48. ES-MS 238(M−1).

Preparation of 3-[(3,4-dimethoxybenzyl)amino]-1-butanesulfonic acid(Compound LM)

To a solution of veratrylamine (1.50 g, 9.0 mmol) in 1,4-dioxane (8 mL)was added 1,4-butane sultone (1.21 g, 8.5 mmol) at room temperature. Themixture was then heated at reflux for 2 hours. The reaction was cooledto room temperature. The solid was collected by filtration, washed withacetone (2×25 mL) and dried on pump. Yield: 18%. ¹H NMR (D₂O, 500 MHz) δ6.96 (m, 3H), 4.04 (s, 2H), 3.74 (m, 6H), 2.95 (t, 2H, J=6.8 Hz), 2.80(t, 2H, J=7.3 Hz), 1.68, (m, 4H). ¹³C (D₂O, 125 MHz) δ ppm 149.19,148.50, 123.82, 123.36, 113.25, 112.17, 55.91, 50.88, 50.24, 46.41,24.55, 21.50. ES-MS 302 (M−1).

Preparation of 4-(adamantan-1-ylamino)-2-hydroxy-1-propanesulfonic acid(Compound KB)

1-adamantaneamine hydrochloride (2.67 g, 14.2 mmol) was treated with 1NNaOH (20 mL) and CH₂Cl₂ (3×20 mL). The organic extracts were combined,dried with Na₂SO₄, filtered, evaporated under reduced pressure and driedin vacuo.

To an 80° C. solution of 1-adamantanamine (2.15 g, 14.2 mmol) in1,4-dioxane (10 mL) and water (5 mL) was added via syringe pump (1 haddition) a solution of 3-chloro-2-hydroxy-propanesulfonic acid, sodiumsalt (1.93 g, 9.7 mmol) in 1,4-dioxane (0.5 mL) and water (10 mL). Thesolution was stirred at reflux overnight. The reaction was cooled toroom temperature. The solvent was evaporated under reduced pressure. Thesolid was suspended in 25% acetone/EtOH. The mixture was heated toreflux for 1 minute. The solid was collected by filtration. The pureproduct crystallized in the filtrate. The product was filtered, washedwith EtOH (2×10 mL), dissolved in water and lyophilized. Yield: 15%. ¹HNMR (D₂O, 500 MHz) δ ppm 4.18 (m, 1H), 3.22 (m, 1H), 3.01 (m, 2H), 2.94(m, 1H), 2.06 (s, 3H), 1.77 (m, 7H), 1.61 (d, 3H), 1.53 (m, 3H). ¹³C(D₂O, 125 MHz) δ ppm 64.23, 57.99, 55.05, 44.10, 38.10, 35.07, 29.03.ES-MS 288 (M−Na (23)).

Preparation of 4-(2-adamantyl)amino-1-butanesulfonic acid (Compound LN)

2-adamantanamine hydrochloride (2.50 g, 13.3 mmol) was treated with 1NNaOH (20 mL) and CH₂Cl₂ (3×20 mL). The organic extracts were combined,dried with Na₂SO₄, filtered, evaporated under reduced pressure and driedin vacuo.

To a solution of 2-adamantanamine (1.06 g, 7.0 mmol) in 1,4-dioxane (6mL) was added 1,4-butane sultone (955 mg, 6.7 mmol). The solution wasstirred at reflux for 5 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration. It was suspended inEtOH (25 mL) and the mixture was heated to reflux for 1 minute beforethe solid was filtered. It was washed with EtOH (1×15 mL) and dried invacuo. Yield: 55%. ¹H NMR (D₂O, 500 MHz) δ ppm 3.29 (m, 1H), 2.97 (m,2H), 2.83 (m, 2H), 2.02 (m, 2H), 1.83 (m, 2H), 1.68 (m, 14H). ¹³C NMR(D₂O, 125 MHz) δ ppm 63.07, 50.25, 45.03, 36.55, 36.31, 29.85, 29.05,26.68, 26.41, 24.47, 21.61. ES-MS 286 (M−1).

Preparation of 3-(2-adamantylamino)-2-hydroxy-1-propanesulfonic acid(Compound KJ)

To an 80° C. solution of 2-adamantanamine hydrochloride (2.50 g, 13.3mmol) and sodium hydroxide (586 mg, 14.6 mmol) in 1,4-dioxane (7 mL) andwater (7 mL) was added via syringe pump (1 hour addition) a solution of3-chloro-2-hydroxy-propane sulfonic acid, sodium salt (1.76 g, 8.9 mmol)in 1,4-dioxane (1 mL) and water (9 mL). The solution was stirred at 80°C. for an additional 4 hours. The reaction was cooled to roomtemperature. The solvent was evaporated under reduced pressure. Thesolid was suspended in EtOH (25 mL). The mixture was heated to refluxfor 1 minute. The solid was removed by filtration. The pure productcrystallized in the filtrate. The product was filtered, washed with EtOH(1×10 mL) and dried in vacuo. Yield: 30%. ¹H NMR (D₂O, 500 MHz) δ ppm4.30 (m, 1H), 3.35 (m, 2H), 3.03 (m, 3H), 2.07 (m, 2H), 1.84 (m, 2H),1.75 (m, 4H), 1.65 (d, 6H). ¹³C (D₂O, 125 MHz) δ ppm 63.31, 55.03,49.51, 36.51, 36.33, 36.26, 29.78, 29.23, 28.81, 26.63, 26.38. ES-MS 289(M+1).

Preparation of3-(bicyclo[2.2.1]hept-2-ylamino)-2-hydroxy-1-propanesulfonic acid(Compound KI)

To an 80° C. solution of exo-2-aminonorbornane (910 mg, 8.2 mmol) andsodium hydroxide (242 mg, 6.1 mmol) in 1,4-dioxane (4 mL) and water (4mL) was added via syringe pump (1 hour addition) a solution of3-chloro-2-hydroxy-propane sulfonic acid, sodium salt (1.09 g, 5.5 mmol)in 1,4-dioxane (0.5 mL) and water (5.5 mL). The solution was stirred at80° C. for an additional 5 hours. The reaction was cooled to roomtemperature. The solvent was evaporated under reduced pressure. Thesolid was suspended in EtOH (25 mL). The mixture was heated to refluxfor 1 minute. The solid was recovered by filtration and it was passedthrough an ion exchange column (Dowex 50×8, 100 g, solvent: water). Theproduct was recrystallized in EtOH/water (99/1). Yield: 17%. ¹H NMR(D₂O, 500 MHz) δ ppm 4.25 (m, 1H), 3.25 (m, 2H), 3.01 (m, 4H), 2.39 (m,1H), 2.27 (m, 1H), 1.69 (m, 1H), 1.51 (m, 1H), 1.38 (m, 3H), 1.19 (m,1H), 1.07 (m, 2H). ¹³C (D₂O, 125 MHz) δ ppm 63.66, 63.50, 62.21, 61.98,54.94, 50.11, 50.04, 39.26, 39.21, 36.02, 35.97, 35.91, 35.80, 34.70,34.61, 27.11, 27.08, 26.50, 26.45. ES-MS 250 (M−1).

Preparation of 4-[(3-methylbutyl)amino]-2-butanesulfonic acid (CompoundKH)

To a hot solution of isoamylamine (2.0 g, 22.9 mmol) in tetrahydrofuran(THF, 11 mL) was added via syringe pump (2 hour addition) a solution of2,4-butane sultone (3.1 g, 21.8 mmol in THF (total of 5 mL)). Thesolution was stirred at reflux for an additional 2 hours. The reactionwas cooled to room temperature. The solid was recovered by filtrationand it was washed with THF (25 mL) and acetone (25 mL). The solid wasdissolved in water (20 mL) and Dowex 50×8 (10 g) was suspended in thesolution. The mixture was stirred for 15 minutes and the resin wasfiltered. The solvent was evaporated under reduced pressure. Yield: 28%.¹H NMR (H₂O, 500 MHz) δ ppm 3.07 (t, 2H, J=7.8 Hz), 2.92 (t, 2H, J=7.8Hz), 2.87 (m, 1H), 2.06 (m, 1H), 1.77 (m, 1H), 1.51 (m, 1H), 1.42 (m,2H), 1.18 (d, 3H, J=6.8 Hz), 0.78 (d, 3H, J=6.3 Hz). ¹³C(H₂O, 125 MHz) δppm 53.21, 46.32, 45.37, 34.36, 28.16, 25.35, 21.51, 14.79. ES-MS 224(M+1).

Preparation of 2-hydroxy-3-[(3-methylbutyl)amino]-1-propane sulfonicacid (Compound KK)

To a 80° C. solution of isoamylamine (2.0 g, 22.9 mmol) in 1,4-dioxane(9 mL) and water (3 mL) was added via syringe pump (1 h addition) asolution of 3-chloro-2-hydroxy-propane sulfonic acid, sodium salt (3.04g, 15.3 mmol) in 1,4-dioxane (9.5 mL) and water (0.5 mL). The solutionwas stirred overnight at 80° C. The solvent was evaporated. The productwas passed through an ion exchange column (Dowex 50×8, 100 g, solvent:water).

It was recrystallized in absolute EtOH and lyophilized. Yield: 27%. ¹HNMR (H₂O, 500 MHz) δ ppm 4.24 (m, 1H), 3.22 (m, 1H), 3.02 (m, 5H), 1.49(m, 3H), 0.79 (d, 3H, J=6.3 Hz). ¹³C(H₂O, 125 MHz) δ ppm 63.54, 54.89,51.53, 46.48, 34.12, 25.46, 21.56, 21.46. ES-MS 226 (M+1).

Preparation of 3-[(dl)-1-Hydroxy-2-pentyl]amino-1-propane sulfonic acid(Compound KL)

To a 80° C. solution of DL-2-amino-1-pentanol (1.0 g, 9.7 mmol) in1,4-dioxane (5 mL) and water (3 mL) was added via syringe pump (1 houraddition) a solution of 3-chloro-2-hydroxy-propane sulfonic acid, sodiumsalt (1.84 g, 9.2 mmol) in 1,4-dioxane (6 mL) and water (0.5 mL). Thesolution was stirred overnight at 80° C. The solvent was evaporated. Theproduct was passed through an ion exchange column (Dowex 50×8, 100 g,solvent: water). The product was dissolved. It was recrystallized inabsolute EtOH and lyophilized. Yield: 27%. ¹H NMR (H₂O, 500 MHz) δ ppm4.26 (m, 1H), 3.77 (m, 1H), 3.32 (m, 1H), 3.24 (m, 1H), 3.03 (m, 3H),1.54 (m, 2H), 1.29 (m, 2H), 0.81 (t, 3H, J=7.3 Hz). ¹³C(H₂O, 125 MHz) δppm 63.69, 63.60, 59.49, 59.38, 58.81, 58.36, 54.98, 48.68, 48.27,29.32, 28.85, 18.40, 18.38, 13.12. ES-MS 242 (M+1).

Preparation of 4-(cyclohexylamino)-1-butanesulfonic acid (Compound LK)

To a solution of cyclohexylamine (2.0 g, 20.2 mmol) in 1,4-dioxane (13mL) was added 1,4-butane sultone (2.61 g, 19.2 mmol). The solution washeated to reflux for 2 hours. The reaction was cooled to roomtemperature. The solid was collected by filtration, washed with acetone(2×20 mL) and dried in vacuo. Yield: 52%. ¹H NMR (D₂O, 500 MHz) δ ppm2.95 (m, 3H), 2.81 (m, 2H), 1.92 (m, 2H), 1.67 (m, 6H), 1.52 (m, 1H),1.18 (m, 4H), 1.02 (m, 1H). ¹³C (D₂O, 125 MHz) δ ppm 57.32, 50.31,44.01, 29.02, 24.84, 24.68, 24.07, 24.55. ES-MS 236 (M+1).

Preparation of 3-[(1-ethyl-1-methylpropyl)amino]-1-propanesulfonic acid(Compound FP)

The flask was closed with a septum and connected to a 20% NaOH scrubberfor the Ritter Reaction. Potassium cyanide (3.25 g, 50 mmol) was addedto acetic acid (13 mL) and the mixture was stirred for 10 min at roomtemperature. A solution of sulfuric acid (7 mL) in acetic acid (6 mL)was added and the resulting suspension was stirred 10 minutes at roomtemperature. The 3-methyl-3-pentanol (5 g, 48.9 mmol) was addeddrop-wise over a 5 minute period. The mixture was stirred at roomtemperature for 4 hours, at which time some chunks of potassium cyanidewere still visible.

Another portion of potassium cyanide (0.6 g, powdered) was added and themixture was stirred for 18 hours at room temperature. The mixture waspurged with nitrogen for 1 h then poured over ice (approx. 50 g). The pHof the solution was adjusted to 9 with the addition of 20% NaOH (use 50%next time to reduce the volume). The layers were separated and theaqueous layer was extracted with ether (1×20 mL). The combined organiclayers were washed with saturated potassium carbonate (1×5 mL) thendried over magnesium sulfate. The ether was evaporated under reducedpressure to afford a yellow oil (4.11 g, 31.8 mmol, 64%). The oil showedto be a mixture of cis and trans formamide but what otherwise pureenough to be used as such. ¹H NMR (500 MHz, DMSO-d6) δ 0.75-0.80 (m,6H), 1.11-1.12 (m, 3H), 1.40-1.54 (m, 2H), 1.66-1.73 (m, 2H), 7.35-7.45(br s and br d, 1H), [7.88 (s) and 8.08 (d, J=11.7 Hz) for 1H); ¹³C NMR(125 MHz, DMSO-d6) δ 7.7, 7.9, 23.4, 24.2, 30.6, 33.8, 55.6, 160.3,163.3

The 1-ethyl-1-methyl-propylformamide (4.00 g, 31.1 mmol) was added to20% NaOH (40 mL). The mixture was heated to reflux for 4 hours then wasleft overnight at room temperature. Toluene (10 mL) was added and thelayers were separated. The organic layer was dried over sodium sulfatethen filtered. The final volume of the filtrate was about 30 mL. It wasused as such in the next step.

A solution of 1,3-propanesultone (2.5 g, 20 mmol) in 2-butanone (10 mL)was added to a solution of 3-methyl-3-ethyl-3-propylamine in toluene(total volume: 30 mL). The mixture was heated to reflux for 5 hours thenwas cooled to room temperature. The solid was collected bysuction-filtration and rinsed with acetone (2×5 mL). The solid was driedovernight at 45° C. in the vacuum oven. The title compound was obtainedas a fine white solid (3.63 g, 16.3 mmol, 33% overall yield). ¹H NMR(500 MHz, DMSO-d6) δ 0.79 (t, J=7.3 Hz, 6H), 1.15 (s, 3H), 1.53-1.59 (m,4H), 1.97-2.00 (m, 2H), 2.89 (t, J=7.1 Hz, 2H), 3.03 (t, J=7.6 Hz, 2H);¹³C NMR (125 MHz, DMSO-d6) δ 6.8, 20.2, 21.9, 27.7, 39.7, 48.2, 63.3ES-MS 224; (M+H)

Preparation of3-({2-hydroxy-1,1-dimethyl-2-(3-methoxyphenyl)ethyl]amino)-1-propanesulfonicacid (Compound NG)

To a cooled solution of sodium methoxide (0.5 M in MeOH, 25 mLl) wasadded via syringe over a 10 minutes period 2-nitropropane (5.0 g, 56mmol). The reaction mixture was stirred at room temperature for 30minutes and recooled before m-anisaldehyde (6.8 mL, 56 mmol) was added.The reaction mixture was stirred at room temperature overnight. Themixture was neutralized with Amberlite IR-120 (strongly acidic). Theresin was removed by filtration and washed with MeOH (2×20 mL). Thefiltrate was evaporated. The resulting oil was purified by flashchromatography: 98% Hexanes/EtOAc to 90% Hexanes/EtOAc, affording thedesired nitro compound (5.70 g, 45%).

To a solution of the nitro compound (5.70 g, 25.3 mmol)) in MeOH (25 mL)was added 6M HCl (25 mL). After cooling to 5° C., zinc powder (8.2 g,125 mmol) was added. The suspension was stirred at 0-5° C. and at roomtemperature overnight. The mixture was filtered on a celite pad. Thefilter cake was washed with MeOH (2×20 mL). The combined filtrates wereevaporated under reduced pressure. The residue was dissolved in EtOAc(40 mL). The mixture was extracted with 5% NaOH (1×40 mL). The aqueousphase was extracted with EtOAc (2×40 mL). The combined organic extractswere dried with Na₂SO₄, filtered, evaporated and dried in vacuo toafford the corresponding amine. The amine (2.15 g, 44%) was used withoutfurther purification.

To a solution of amine (2.15 g, 11.0 mmol) in Pinacolone (6 mL) andtoluene (6 mL) was added 1,3-propane sultone (1.28 g, 10.5 mmol). Thesolution was stirred at reflux overnight. The reaction mixture wascooled to room temperature. The solid material was collected byfiltration, was washed with acetone (2×20 mL). The solid was suspendedin EtOH (30 mL). The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature. The white solid was filtered,washed with acetone (2×15 mL) and dried in a vacuum oven at 50° C.,affording the title compound, 2.26 g (66%). ¹H NMR (DMSO, 500 MHz) δ ppm8.45 (s (broad), 1H), 7.26 (t, 1H, J=7.9 Hz), 6.89 (m, 3H), 6.30 (d, 1H,J=3.2 Hz), 4.69 (d, 1H, J=3.8 Hz), 3.74 (s, 3H), 3.10 (m, 2H), 2.62 (t,2H, J=6.7 Hz), 2.00 (m, 2H), 1.07 (m, 6H). ¹³C (DMSO, 125 MHz) δ ppm159.24, 142.09, 129.45, 120.80, 114.30, 113.76, 74.16, 62.48, 55.92,50.10, 41.57, 23.30, 21.18, 19.37. ES-MS 316 (M−1).

Preparation of3-{[1-(4-methylbenzyl)cyclohexyl]amino}-1-propane-1-sulfonic acid(Compound NH)

NaOMe (0.5M, 40 mL) was added to nitrocyclohexane (2.58 g, 20 mmol) andthe solution was stirred for 30 minutes then concentrated to afford awhite solid. To this solid was added 4-methylbenzylpyrridinium (6.6 g,13 mmol) and DMSO (20 mL). The mixture was heated at 100° C. for 15hours then cooled to rt and diluted with HCl (1M) and EtOAc. Afterseparation of the two phases, the organic layer was washed twice withHCl (1M) then concentrated to obtain an oily crude. Methanol was addedto precipitate the pyridinium byproduct which was filtered off, and thefiltrate was concentrated and purified by column using Hex:EtOAc 90:10to obtain the desired nitro (still contaminated with the pyridiniumsalt). 2 g, 66% yield.

To a stirred solution of the nitro (2.0 g, 8.58 mmol) in methanol (20mL) was added a spatula of Raney-Ni in water. The suspension washydrogenated under atmospheric pressure of hydrogen for 15 hours (TLCindicates complete consumption of the starting material) then filteredon celite and concentrated under reduced pressure. The crude waspurified by column using CH₂Cl₂:MeOH 80:10 to afford 1.2 g of thecorresponding amine.

To a stirred solution of the amine (800 mg, 3.93 mmol) in THF (8 mL) wasadded 1,3-propane sultone (480 mg, 3.93 mmol). The reaction mixture wasstirred at reflux for 15 hours then cooled to room temperature. Thesolid was collected by filtration and was washed with THF. The solid wassuspended in EtOH (10 mL) and stirred at reflux for 1 hour. Thesuspension was then cooled to room temperature. The solid was collectedby filtration, washed with ethanol and dried under high vacuum to affordthe title compound, 1.1 g (86%). ¹H NMR (500 MHz, DMSO-d₆) δ 1.18-1.78(m, 10H), 2.00 (m, 2H), 2.29 (s, 3H), 2.65 (m, 2H), 2.92 (s, 2H), 3.12(m, 2H), 7.10-7.16 (m, 2H), 8.39 (bs, 2H). ¹³NMR (125 MHz, DMSO-d₆) δ20.74, 21.35, 22.80, 25.05, 31.60, 41.04, 50.24, 61.63, 129.71, 131.38,132.44, 136.80. ES-MS 324 (M−1).

Preparation of3-{[2-(4-methoxyphenyl)-1,1-dimethylethyl]amino}-1-propanesulfonic acid(Compound NI)

To a stirred solution of the phenol (233 mg, 1 mmol) in DMF/THF (2.5mL/2.5 mL) was added MeI (93 uL, 1.5 mmol) followed by K₂CO₃ (276 mg, 2mmol). The suspension was heated at reflux for 15 hours then dilutedwith HCl (1M) and with EtOAc. The organic layer was washed with HCl(1M)then concentrated under high vacuum. The crude was purified by columnusing Hex:EtOAc 90:10 to obtain 215 mg of the methoxy (87% yield).

To a stirred solution of the nitro (300 mg, 1.2 mmol) in methanol (5 mL)was added asmall spatula of Raney-Ni in water. The suspension washydrogenated under atmospheric pressure of hydrogen for 3 hours (TLCindicates complete consumption of the starting material) then filteredon prewashed celite and concentrated under reduced pressure. The crudeamine was used as such in the next step.

To the crude amine (240 mg, 1.34 mmol) in solution in THF (3 mL) wasadded 1,3-sultone (181 mg, 1.48 mmol) and the mixture was heated atreflux of THF for 12 hours. The suspension of was cooled down andfiltered. The solid was dried to afford 270 mg of the homotaurin as awhite solid (67% yield). ¹H NMR (500 MHz, D₂O) δ 1.11 (s, 6H), 2.00 (m,2H), 2.67 (m, 2H), 2.80 (m, 2H), 3.12 (m, 2H), 3.74 (s, 3H), 6.90 (m,2H), 7.14 (m, 2H), 8.61 (bs, 2H). ES-MS 272 (M−1). ES-MS 300 (M−1).

Preparation of3-{[2-hydroxy-1,1-dimethyl-2-(4-methylphenyl)ethyl]amino}-1-propanesulfonicacid (Compound NJ)

To a solution of 2-nitropropane (3.0 g, 34 mmol), p-tolualdehyde (4.0mL, 34 mmol) and Tetrahydrofuran (30 mL) was added Amberlyst A-21 (7 g).The reaction mixture was stirred at room temperature for 40 hours. Theresin was removed by filtration and washed with THF (2×20 mL). Thefiltrate was evaporated. The resulting oil was purified by flashchromatography: 98% Hexanes/EtOAc to 90% Hexanes/EtOAc, affording thedesired nitro compound (820 mg, 12%).

A suspension of Pd/C and the nitro compound (820 mg, 3.9 mmol) in EtOAc(10 mL) was stirred under H₂ (1 atm) overnight. The mixture was filteredon a celite pad. The celite was washed with EtOAc (2×15 mL). Thecombined filtrates were evaporated under reduced pressure to afford thecorresponding amine. The amine (470 mg, 67%) was used without furtherpurification.

To a solution of amine (470 mg, 2.6 mmol) in pinacolone (5 mL) andToluene (5 mL) was added 1,3-propane sultone (310 mg, 2.5 mmol). Thesolution was stirred at reflux for 4 hours. The reaction mixture wascooled to room temperature. The solid was collected by filtration, waswashed with acetone (2×10 mL) and dried in vacuo, affording the titlecompound, 196 mg (26%). ¹H NMR (DMSO, 500 MHz) δ ppm 8.46 (s (broad),1H), 7.24 (d, 2H, J=7.8 Hz), 7.16 (d, 2H, J=8.3 Hz), 6.23 (d, 1H, J=3.9Hz), 4.68 (d, 1H, J=3.9 Hz), 3.11 (m, 2H), 2.63 (t, 2H, J=6.8 Hz), 2.29(s, 3H), 2.00 (m, 2H), 1.04 (s, 6H). ¹³C (DMSO, 125 MHz) δ ppm 137.69,137.56, 129.03, 128.45, 74.07, 62.38, 49.91, 41.35, 22.99, 21.39, 20.81,18.76. ES-MS 300 (M−1).

Preparation of3-{[1,1-dimethyl-2-(4-methylphenyl)ethyl]amino}-1-propanesulfonic acid(Compound NK)

NaOMe (0.5M, 20 mL) was added to 2-nitropropane (890 mg, 10 mmol) andthe solution was stirred for 30 minutes then concentrated to afford awhite solid. To this solid was added 4-methylbenzylpyrridinium (3.3 g,15 mmol) and DMSO (15 mL). The mixture was heated at 100° C. for 15hours then cooled to room temperature and diluted with HCl (1M) andEtOAc. After separation of the two phases, the organic layer was washedtwice with HCl (1M) then concentrated to obtain an oily crude product.Methanol was added to precipitate the pyridinium byproduct which wasfiltered off, and the filtrate was concentrated and purified by columnusing Hex:EtOAc 90:10 to obtain the desired nitro but still contaminatedwith the pyridinium salt. 1.32 g, 66% yield.

To a stirred solution of the nitro (700 mg, 3.62 mmol) in methanol (10mL) was added a small spatula of Raney-Ni in water. The suspension washydrogenated under atmospheric pressure of hydrogen for 15 hours (TLCindicates complete consumption of the starting material) then filteredon celite and concentrated under reduced pressure. The crude amine wasused as such in the next step.

To a stirred solution of the amine (550 mg, 3.39 mmol) in THF (8 mL) wasadded 1,3-propane sultone (414 mg, 3.39 mmol). The reaction mixture wasstirred at reflux for 6 hours then cooled to room temperature. The solidwas collected by filtration and was washed with THF. The solid wassuspended in EtOH (5 mL) and stirred at reflux for 1 hour. Thesuspension was then cooled to room temperature. The solid was collectedby filtration, washed with ethanol and dried under high vacuum to affordthe title compound, 210 mg (22%). ¹H NMR (500 MHz, DMSO-d₆) δ 1.13 (s,6H), 2.00 (m, 2H), 2.66 (dd, J=7.0 & 7.0 Hz, 2H), 2.75 (s, 2H), 3.10(dd, J=7.0 & 7.0 Hz, 2H), 6.72 (d, J=8.3 Hz, 2H), 7.00 (d, J=8.3 Hz,2H), 8.60 (bs, 2H), 9.36 (s, 1H). ¹³NMR (125 MHz, DMSO-d₆) δ 23.1, 41.2,43.2, 49.8, 59.4, 115.7, 125.8, 132.3, 157.1. ES-MS 284 (M−1).

Preparation of 4-(bicyclo[2.2.1]hept-2-ylamino)-1-butanesulfonic acid(Compound MX)

To a solution of exo-2-aminonorbornane (800 mg, 7.2 mmol) in 1,4-dioxane(5 mL) was added 1,4-butanesultone (1.00 g, 7.0 mmol) at roomtemperature. The solution was stirred at reflux for 5 hours. Thereaction was cooled to room temperature. The solid was collected byfiltration, washed with 1,4-dioxane (2×20 mL) and dried in vacuo. Thesolid was suspended in EtOH (20 mL). The mixture stirred at reflux for 5minutes before the solid was filtered and it was dried in vacuo. Yield:51%. ¹H NMR (D₂O, 500 MHz) δ ppm, 3.00 (m, 1H), 2.95 (m, 2H), 2.81 (m,2H), 2.34 (m, 1H), 2.26 (m, 1H), 1.68 (m, 5H), 1.48 (m, 1H), 1.37 (m,3H), 1.18 (d, 1H, J=10.7 Hz), 1.06 (m, 2H). ¹³C (D₂O, 125 MHz) δ ppm.61.82, 50.26, 45.44, 39.37, 35.93, 35.78, 34.67, 27.11, 26.40, 24.63,21.56. ES-MS 248 (M+1).

Preparation of 4-(1H-benzimidazol-2-ylthio)-2-butanesulfonic acid(Compound NE)

To a hot solution of 2-mercaptobenzimidazole (2.0 g, 13.3 mmol) in1,4-dioxane (12 mL) and water (3 mL) was added via syringe pump (1 houraddition) a solution of 2,4-butane sultone (1.80 g, 12.7 mmol in1,4-dioxane (total of 3 mL)). The solution was stirred at reflux for anadditional 3 hours. The solid was collected by filtration. It was washedwith acetone (2×20 mL) and dried in vacuo. Yield: 86%. ¹H NMR (DMSO, 500MHz) δ ppm 7.64 (m, 2H), 7.44 (m, 2H), 3.63 (t, 2H, J=7.3 Hz), 2.68 (m,1H), 2.10 (m, 1H), 1.90 (m, 1H), 1.15 (d, 3H, J=6.8 Hz). ¹³C (DMSO, 125MHz) δ ppm 152.64, 133.32, 125.66, 113.67, 53, 10, 39.72, 33.48, 30.14,16.85. ES-MS 287 (M+1).

Preparation of 3-[(1,1-diethylpropyl)amino]-1-propanesulfonic acid(Compound NM)

For the Ritter reaction, the flask was closed with a septum andconnected to a 20% NaOH scrubber. Potassium cyanide (powdered, 6.19 g,95 mmol) was added to acetic acid (28 mL) in portions over a 2 minuteperiod. The mixture was stirred for 10 minutes at room temperature. Asolution of sulfuric acid (14 mL) in acetic acid (11 mL) was added overa 2 minute period and the resulting suspension was stirred 10 minutes atroom temperature. The 3-ethyl-3-pentanol (10 g, 86 mmol) was addeddropwise over a 12 minute period. The mixture was stirred at roomtemperature for 22 hours and then poured over ice (approx. 100 g). ThepH of the solution was adjusted to 9 with the addition of 50% NaOH(about 120 g). The layers were separated and the aqueous layer wasextracted with ether (1×50 mL). The combined organic layers were washedwith saturated sodium carbonate (1×5 mL) then dried over sodium sulfate.The ether was evaporated under reduced pressure to afford a beige waxysolid (12.31 g, 84.76 mmol, 99%). The solid showed to be a mixture ofcis and trans formamide but what otherwise pure enough to be used assuch. ¹H NMR (500 MHz, CDCl₃) δ [0.81 (t, J=7.3 Hz) and 0.86 (t, J=7.3Hz) for 9H], [1.55 (q, J=7.3 Hz) and 1.70 (q, J=7.3 Hz) for 9H], [4.83and 5.65 (br s, 1H)], [8.09 (s) and 8.16 (d, J=12.7 Hz) for 1H]; ¹³C NMR(125 MHz, CDCl₃) δ 7.2, 26.7, 29.5, 58.3, 60.0, 160.2, 163.4

A solution of NaOH (20%, 80 mL) was added to a solution of the1,1-diethyl-1-propylformamide (12.31 g, 84.8 mmol) in toluene (10 mL).The hydrolysis was not completed after 4 hours at reflux. A catalyticamount of Triton X-100 and 1,4-dioxane (2 mL) were added. The mixturewas heated to reflux for 48 hours then the mixture was cooled at roomtemperature. Some sodium chloride (10 g) was added to facilitate thephase separation. The layers were separated and the aqueous layer wasextracted with toluene (1×20 mL). The combined organic layers werewashed with brine (1×10 mL) then dried over sodium sulfate and filtered.The final volume of the filtrate was about 40 mL. It was used as such inthe next step.

A solution of 1,3-propanesultone (8.4 g, 68 mmol) in 2-butanone (20 mL)was added dropwise over a 10 minute period to a solution of1,1-diethyl-3-propylamine in toluene (40 mL total). The mixture washeated to reflux for 5 hours then was cooled to room temperature. Thesolid was collected by suction-filtration and rinsed with acetone (2×5mL). The solid was dried overnight at 45° C. in the vacuum oven. Thetitle compound was obtained as a fine white solid (13.33 g, 56.16 mmol,65% overall yield). ¹H NMR (500 MHz, D₂O) δ 0.81 (m, 9H), 1.59 (m, 6H),2.04 (m, 2H), 2.94 (m, 2H), 3.05 (m, 2H); ¹³C NMR (125 MHz, D₂O) δ 6.5,21.9, 25.1, 39.6, 48.3, 66.1; ES-MS 238 (M+H)

Preparation of 3-[(1-ethylcyclopentyl)amino]-1-propanesulfonic acid(Compound NN)

For the Ritter reaction, the flask was closed with a septum andconnected to a 20% NaOH scrubber. Sodium cyanide (powdered, 1.07 g, 22mmol) was added in one portion to acetic acid (5 mL). The mixture wasstirred for 5 minutes at room temperature. A solution of sulfuric acid(3 mL) in acetic acid (3 mL) was added dropwise over a 2 minute period.The suspension was stirred 10 minutes at room temperature then asolution of 1-ethyl-1-cyclopentanol (2 g, 17.5 mmol) in acetic acid (1mL) was added dropwise over a 2 minute period. The mixture was stirredat room temperature for 22 hours then poured on ice (about 50 g). The pHof the solution was adjusted to 9 with the addition of 50% NaOH (about24 g). The layers were separated and the aqueous layer was extractedwith ether (2×10 mL). The combined organic layers were washed withsaturated sodium carbonate (1×5 mL) then dried over magnesium sulfate.The ether was evaporated under reduced pressure to afford a clear yellowoil (2.35 g, 16.6 mmol, 95%). The oil showed to be a mixture of cis andtrans formamide but what otherwise pure enough to be used as such. ¹HNMR (300 MHz, CDCl₃) δ 0.85-0.96 (m, 3H), 1.61-1.97 (m, 10H), [5.23 and5.77 (br s, 1H)], [8.06 (s) and 8.17 (d, J=12.3 Hz) for 1H]; ¹³C NMR (75MHz, CDCl₃) δ 9.0, 9.3, 23.0, 23.8, 30.1, 34.0, 37.8, 38.6, 160.3, 163.4

A solution of NaOH (20%, 20 mL) was added to a mixture of the crude1-ethyl-1-cyclopentylformamide (2.3 g), Triton X-100 (2 drops) andtetrabutylammonium bromide (45 mg). The mixture was heated to reflux for3 days then cooled at room temperature. Some sodium chloride (5 g) wasadded to facilitate the phase separation. The layers were separated andthe aqueous layer was extracted with MTBK (2×10 mL) and toluene (1×2mL). The combined organic layers were washed with brine (1×5 mL) thendried over sodium sulfate and filtered. The filtrate was used as such inthe next step. ¹H NMR (500 MHz, CD₃OD) δ 0.95 (t, J=7.3 Hz, 3H)1.15-1.65 (m, 8H), 1.76-1.80 (m, 2H); ¹³C NMR (125 MHz, CD₃OD) δ 9.3,25.2, 36.7, 40.6, 62.4

A solution of 1,3-propane sultone (1.9 g, 15 mmol) in toluene (2 mL) wasadded to the crude solution of 1-ethyl-1-cyclopentylamine inMTBK/toluene (total volume 30 mL). The mixture was heated to reflux for20 hours then cooled to room temperature. The solid was collected bysuction filtration, rinsed with acetone (2×5 mL). There were specs inthe dry crude solid (3.15 g). The solid was recrystallized in hot 90%ethanol. The solid was dried overnight at 60° C. in the vacuum oven. Thetitle compound was obtained as a fine white solid (2.35 g, 10 mmol, 57%overall yield). ¹H NMR (500 MHz, D₂O) δ 0.81 (t, J=7.3 Hz, 3H),1.53-1.67 (m, 8H), 1.71-1.77 (m, 2H), 1.98 (q, J=7.6 Hz, 2H), 2.88 (t,J=7.8 Hz, 2H), 3.04 (t, J=7.3 Hz, 2H); ¹³C NMR (125 MHz, D₂O) 7.3, 22.0,24.3, 29.3, 34.6, 41.1, 48.2, 70.4; ES-MS 234 (M−H)

Preparation of 3-[(1-ethylcycloheptyl)amino]-1-propanesulfonic acid(Compound NO)

A solution of cycloheptanone (5.69 mL, 50 mmol) was added dropwise to acold (0° C.) solution of 1-Methyl magnesium bromide in THF (50 mL). Themixture was stirred 1.5 hours at room temperature then for 1 hour atreflux. The reaction mixture was cooled with an ice-water bath and thereaction was quenched with the addition of saturated ammonium chloride(20 mL). The layers were separated and the aqueous phase was extractedonce with ether (1×20 mL). The combined organic layers were dried overmagnesium sulfate and the ether was removed under reduced pressure. Thecrude oil (6.34 g) was distilled to afford a clear oil (3.81 g) thatcontained some cycloheptanone. The product was placed under high vacuumuntil the cycloheptanone contains was less than 5 mol % (2.81 g, 19.8mmol, 39%). ¹H NMR (300 MHz, CDCl₃) δ 0.91 (t, J=12.3 Hz, 3H), 1.19 (brs, 1H), 1.36-1.68 (m, 14H); ¹³C NMR (75 MHz, CDCl₃) δ 7.9, 22.6, 30.0,35.9, 40.8, 75.5

For the Ritter reaction, the flask was closed with a septum andconnected to a 20% NaOH scrubber. Sodium cyanide (powdered, 1.20 g, 24mmol) was added in one portion to acetic acid (5 mL). The mixture wasstirred for 5 minutes at room temperature. A solution of sulfuric acid(3.5 mL) in acetic acid (4 mL) was added dropwise over a 5 minuteperiod. The suspension was stirred 10 minutes at room temperature then asolution of 1-ethyl-1-cycloheptanol (2 g, 17.5 mmol) in acetic acid 31mL) was added dropwise over a 5 minute period. The mixture was stirredat room temperature for 22 hours then poured on ice (about 50 g). The pHof the solution was adjusted to 9 with the addition of 50% NaOH (about24 g). The layers were separated and the aqueous layer was extractedwith ether (2×10 mL). The combined organic layers were washed withsaturated sodium carbonate (1×5 mL) then dried over magnesium sulfate.The ether was evaporated under reduced pressure to afford a clear yellowoil (3.18 g, 18.8 mmol, 95%). A trace the cycloheptanone was stillpresent has indicated by the proton NMR. The oil showed to be a mixtureof cis and trans formamide but what otherwise pure enough to be used assuch. ¹H NMR (300 MHz, CDCl₃) δ 0.80-0.91 (m, 3H), 1.49-1.96 (m, 14H),[5.07 and 5.70 (br s, 1H)], [8.07 (d, J=2.1 Hz) and 8.17 (d, J=12.3 Hz)for 1H]; ¹³C NMR (75 MHz, CDCl₃) δ 7.9, 8.2, 22.3, 22.5, 29.6, 29.7,31.2, 35.7, 38.3, 39.9, 58.7, 60.4, 160.1, 163.3

A solution of NaOH (20%, 20 mL) was added to a mixture of the crude1-ethyl-1-cycloheptylformamide (3.18 g), Triton X-100 (2 drops) andtetrabutylammonium bromide (45 mg). The mixture was heated to reflux for4 days then cooled at room temperature. The layers were separated andthe aqueous layer was extracted with ether (2×5 mL). The etherealsolution was dried over magnesium sulfate and filtered. Then, a solutionof 2N HCl was added and the mixture was concentrated to a thick oil. Theoil was diluted with 1N HCl and washed with ether (2×10 mL). The pH ofthe aqueous layer was adjusted to 10 with the addition of 50% NaOH. Theorganic layer was separated and the aqueous phase was extracted withMTBK (2×4 mL). The combined organic layers were washed with brine (1×5mL) then dried over sodium sulfate and filtered. The filtrate was usedas such in the next step. A trace of cycloheptanone and1-ethyl-1-cycloheptylformamide were still visible on the proton NMR. ¹HNMR (500 MHz, CD₃OD) δ 0.90 (t, J=7.2 Hz, 3H) 1.42-1.64 (m, 14H); ¹³CNMR (125 MHz, CD₃OD) δ 8.1, 23.9, 31.6, 36.6, 42.1, 55.7

A solution of 1,3-propane sultone (1.0 g, 8 mmol) in toluene (3 mL) wasadded to the crude solution of 1-ethyl-1-cycloheptylamine inMTBK/toluene (total volume 20 mL). The mixture was heated to reflux for1 hour, for the night at room temperature then another hour at reflux.The mixture was cooled to room temperature. The solid was collected bysuction filtration, rinsed with acetone (2×5 mL) then ethanol (1×5 mL).The solid was dried 5 hours at 60° C. in the vacuum oven. The titlecompound was obtained as a fine white solid (1.35 g, 5.12 mmol, 10%overall yield). ¹H NMR (500 MHz, DMSO-d6) δ 0.87 (t, J=7.6 Hz, 3H),1.37-1.41 (m, 2H), 1.49 (br s, 4H), 1.58-1.77 (m, 8H), 2.00 (q, J=6.2Hz, 2H), 2.68 (t, J=6.3 Hz, 2H), 2.99 (br s, 2H), 8.42 (br s, 2H); ¹³CNMR (125 MHz, DMSO-d6) 7.1, 21.6, 21.9, 28.6, 29.7, 34.8, 40.8, 49.9,64.1; ES-MS 262 (M−H)

Preparation of 3-[(1,3-dimethylbutyl)amino]-1-propanesulfonic acid(Compound NP)

A solution of 1,3-propanesultone (6.1 g, 50 mmol) in a mixture ofMTBK/toluene (60:40, 15 mL) was added in one portion to a solution of1,3-dimethylbutylamine (5 g, 49 mmol) in mixture of MTBK/toluene (60:40,25 mL). The mixture was heated under reflux for 3 hours then at roomtemperature for the night. Ethanol (5 mL) was added and the mixture washeated at reflux for 1 hours then it was cooled to 4° C. The solid wascollected by suction filtration, rinsed with acetone (3×10 mL). Thesolid was dried overnight at 60° C. in the vacuum oven. The titlecompound was obtained as a fine white solid (9.33 g, 41.8 mmol, 85%overall yield). ¹H NMR (500 MHz, D₂O) δ 0.75 (d, J=6.8 Hz, 3H), 0.81 (d,J=6.8 Hz, 3H), 1.17 (d, J=6.3 Hz, 3H), 1.55-1.60 (m, 1H), 1.95-2.02 (m,2H), 2.88 (t, J=7.3 Hz, 2H), 3.03-3.11 (m, 2H), 3.21-3.25 (m, 1H); ¹³CNMR (125 MHz, D₂O) 15.7, 50.6, 21.7, 22.7, 24.3, 41.7, 43.3, 48.1, 53.4;ES-MS 222 (M−H)

Preparation of 3-{[(1S)-1,2,2-trimethylpropyl]amino}-1-propanesulfonicacid (Compound NQ)

A solution of 1,3-propane sultone (6.6 g, 54 mmol) in toluene (20 mL)was added in one portion to a solution of (S)-3,3-Dimethyl-2-butylamine(5 g, 49 mmol) in MTBK (20 mL). The mixture was heated to reflux. Within1 hour, it had turned to a lump. More solvent (10 mL of MTBK, 5 mL oftoluene then 4 mL of ethanol) were added to restore the stirring. Themixture was then heated at reflux for 18 hours and then it was cooled to3° C. The solid was collected by suction filtration, rinsed with acetone(2×10 mL). The solid was dried overnight at 60° C. in the vacuum oven.The title compound was obtained as a fine white solid (9.79 g, 43.8mmol, 89% overall yield). ¹H NMR (300 MHz, D₂O) δ 0.88 (s, 9H), 1.15 (d,J=6.7 Hz, 3H), 1.94-2.14 (m, 2H), 2.87-3.10 (m, 4H), 3.18-3.28 (m, 1H);¹³C NMR (125 MHz, D₂O) 11.2, 21.2, 25.2, 33.2, 45.5, 48.3, 64.1; ES-MS222 (M−H)

Preparation of 3-[(1-ethylcyclohexyl)amino]-1-propanesulfonic acid(Compound NR)

For the Ritter reaction, the flask was closed with a septum andconnected to a 20% NaOH scrubber. Potassium cyanide (powdered, 3.0 g, 46mmol) was added in portions to acetic acid (10 mL). The mixture wasstirred for 10 minutes at room temperature. A solution of sulfuric acid(6 mL) in acetic acid (5 mL) was added dropwise over a 10 minute period.The suspension was stirred for 10 minutes at room temperature then the1-methyl-1-cycloheptanol (5 g, 39.0 mmol) was added dropwise. Themixture was stirred at room temperature for 22 hours then poured on ice(about 50 g). The pH of the solution was adjusted to 9 with the additionof 50% NaOH (about 70 g). The layers were separated and the aqueouslayer was extracted with ether (2×20 mL). The combined organic layerswere washed with saturated sodium carbonate (1×5 mL) then dried oversodium sulfate. The ether was evaporated under reduced pressure toafford a clear yellow oil (5.44 g, 90%). The oil showed to be a mixtureof cis and trans formamide but what otherwise pure enough to be used assuch. ¹H NMR (400 MHz, CDCl₃) δ 0.81-0.90 (m, 3H), 1.19-1.60 (m, 9H),1.68-1.71 (m, 1H), 1.78-1.84 (m, 1H), 2.02-2.06 (m, 1H), [5.03 and 5.65(br s, 1H)], [8.15 (s) and 8.18 (d, J=12.4 Hz) for 1H]; ¹³C NMR (100MHz, CDCl₃) δ 7.2, 7.6, 21.5, 21.9, 25.8, 25.9, 31.0, 34.7, 35.1, 36.4,55.0, 56.9, 160.7, 163.8

A solution of NaOH (20%, 40 mL) was added to the crude1-ethyl-1-cyclohexylformamide (5.44 g). The mixture was heated to refluxfor 3 hours. The reaction was not completed by proton NMR. It was heatedto reflux overnight. The reaction was still not completed. A catalyticamount of tetrabutylammonium bromide (200 mg) was added. The mixture washeated to reflux for 3 more days then cooled at room temperature. Thelayers were separated and the aqueous layer was extracted with MTBK(2×10 mL). The combined organic layers were washed with brine (1×5 mL),then dried over sodium sulfate and filtered. The filtrate was used assuch in the next step. ¹H NMR (400 MHz, CD₃OD) 0.84-0.88 (m, 3H),1.30-1.55 (m, 12H); ¹³C NMR (100 MHz, CD₃OD) δ 6.2, 22.0, 25.8, 37.4,50.3, 66.9

A solution of 1,3-propane sultone (3.2 g, 26 mmol) in toluene (6 mL) wasadded to the crude solution of 1-ethyl-1-cyclohexylamine in MTBK (totalvolume: 30 mL). The mixture was heated to reflux for 18 hours. Anotherportion of 1,3-propane sultone (0.7 g) in toluene (6 mL) was added. Themixture was heated to reflux for another 4 hours then it was cooled toroom temperature. The solid was collected by suction filtration, rinsedwith acetone (2×5 mL). The solid was dried overnight at 60° C. in thevacuum oven. The title compound was obtained as a fine white solid (3.55g, 14.2 mmol, 36% overall yield). ¹H NMR (500 MHz, D₂O) δ 0.84 (t, J=7.3Hz, 3H), 1.18-1.24 (m, 1H), 1.39-1.49 (m, 4H), 1.54-1.60 (m, 3H),1.73-1.81 (m, 4H), 2.06 (q, J=7.6 Hz, 2H), 2.96 (t, J=7.8 Hz, 2H), 3.10(t, J=7.3 Hz, 2H)); ¹³C NMR (100 MHz, D₂O) 6.1, 21.2, 22.0, 23.7, 24.6,32.0, 39.1, 48.3, 62.9; ES-MS 248 (M−H)

Preparation of3-{[1-(4-hydroxyphenyl)-2-methyl]-2-propylamine}-1-propanesulfonic acid(Compound NS)

Benzyl alcohol (1.2 g, 10 mmol), the tetrabutylammonium fluoride (5 mL,5 mmol) and 2-nitropropane (1.78 g, 20 mmol) were placed in a sealedtube and heated at 130° C. for 15 hours. The reaction was cooled anddiluted with EtOAc. The resulting solution was washed with water, driedand concentrated to yield a dark oil. Chromatography over silica elutingwith Hex:EA 70:30 gave a yellowish solid 1.42 g, 73%.

To a stirred solution of the nitro (800 mg, 4.12 mmol) in methanol (20mL) was added a small spatula of Raney-Ni in water. The suspension washydrogenated under atmospheric pressure of hydrogen for 15 hours (TLCindicates complete consumption of the starting material) then filteredon celite and concentrated under reduced pressure. The correspondingamine was used as such in the next step.

To a stirred solution of the amine (750 mg, 4.57 mmol) in THF (9 mL) wasadded 1,3-propane sultone (614 mg, 5.02 mmol). The reaction mixture wasstirred at reflux for 4 hours then cooled to room temperature. The solidwas collected by filtration and was washed with THF. The solid wassuspended in EtOH (10 mL) and stirred at reflux for 1 hour. Thesuspension was then cooled to room temperature. The solid was collectedby filtration, washed with ethanol and dried under high vacuum to affordthe title compound, 1.1 g (85%). ¹H NMR (500 MHz, DMSO-d₆) δ 1.21 (s,6H), 1.42-1.48 (m, 2H), 1.55-1.50 (m, 2H), 1.90-2.00 (m, 2H), 2.68 (dd,J=7.0 & 7.0 Hz, 2H), 3.00 (dd, J=7.0 & 7.0 Hz, 2H), 3.40 (dd, J=6.2 Hz,2H). ¹³C NMR (125 MHz, DMSO-d₆) δ 23.0, 23.6, 27.1, 35.1, 41.1, 50.0,58.8, 61.3. ES-MS 286 (M−1).

Preparation of 3-[(5-hydroxy-1,1-dimethylbutyl)amino]-1-propanesulfonicacid (Compound NT)

A mixture of the acrylate (2.7 mL, 30 mmol), nitropropane (5.4 mL, 60mmol) and NaOMe (0.5 M, 12 mL) was stirred for 15 hours. The reactionmixture was quenched with HCl (1M) and extracted with EtOAc. The organiclayer was dried and concentrated under reduced pressure. The crude waspurified by column using Hex:EtOAc 80:20 to afford the desired product.4.5 g (85% yield)

To a stirred solution of the nitro-ester (1.7 g, 10 mmol) in MeOH/THF 50mL/5 mL was added at −10° C. in one portion LiBH₄ (436 mg, 20 mmol). Thereaction was stirred for 2 h then another portion of LiBH₄ (436 mg, 20mmol) was added. The cooling bath was removed allowing the reaction toreach the room temperature and the stirring was continued for 6 hours.The reaction was acidified with HCl (1M) and concentrated under reducedpressure to remove methanol. The reaction was extracted with EtOAc(3×100 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated. The crude was purified by column on silica using Hex:EA80:20 to 50:50 to afford the desired product (1 g, 70%) along withstarting material (340 mg, 20%).

To a stirred solution of the nitro (1.47 g, 10 mmol) in methanol (20 mL)was added a spatula of Raney-Ni in water. The suspension washydrogenated under atmospheric pressure of hydrogen for 3 hours (TLCindicates complete consumption of the starting material) then filteredon celite and concentrated under reduced pressure. The correspondingamine was used as such in the next step.

To a stirred solution of the amine (600 mg, 5.12 mmol) in THF (10 mL)was added 1,3-propane sultone (626 mg, 5.12 mmol). The reaction mixturewas stirred at reflux for 4 hours then cooled to room temperature. Thesolid was collected by filtration and was washed with THF. The solid wassuspended in EtOH (10 mL) and stirred at reflux for 1 hour. Thesuspension was then cooled to room temperature. The solid was collectedby filtration, washed with ethanol and dried underhigh vacuum to affordthe title compound, 770 mg (87%). ¹H NMR (500 MHz, DMSO-d₆) δ 1.13 (s,6H), 2.00 (m, 2H), 2.66 (dd, J=7.0 & 7.0 Hz, 2H), 2.75 (s, 2H), 3.10(dd, J=7.0 & 7.0 Hz, 2H), 6.72 (d, J=8.3 Hz, 2H), 7.00 (d, J=8.3 Hz,2H), 8.60 (bs, 2H), 9.36 (s, 1H). ¹³NMR (125 MHz, DMSO-d₆) δ 23.1, 41.2,43.2, 49.8, 59.4, 115.7, 125.8, 132.3, 157.1. ES-MS 238 (M−1).

Preparation of 3-{[(1S)-1-(4-chlorophenyl)ethyl]amino}-1-propanesulfonicacid (Compound NU)

To a solution of (1S)-(−)-1-(4-chlorophenyl)ethylamine (5.0 g, 32.1mmol) in Pinacolone (20 mL) and toluene (20 mL) was added 1,3-propanesultone (3.7 g, 30.6 mmol). The solution was stirred at reflux for 4hours. The reaction mixture was cooled to room temperature. The solidwas collected by filtration and was washed with acetone (2×25 mL). Thesolid was suspended in EtOH (60 mL). The suspension was stirred atreflux for 1 hour. The mixture was cooled to room temperature, the solidwas collected by filtration, washed with acetone (2×25 mL) and dried ina vacuum oven at 50° C., affording the title compound, 7.14 g (84%). ¹HNMR (D₂O, 500 MHz) δ ppm 7.39 (dd, 2H, J=2.4 Hz, 6.6 Hz), 7.32 (dd, 2H,J=2.4 Hz, 6.6 Hz), 4.30 (q, 1H, J=6.8 Hz), 3.00 (m, 1H), 2.84 (m, 1H),2.79 (t, 2H, J=7.3 Hz), 1.94 (m, 2H), 1.53 (d, 3H, J=6.8 Hz). ¹³C (D₂O,125 MHz) δ ppm 135.16, 134.43, 129.56, 129.36, 57.93, 48.03, 44.45,21.50, 18.12. [α]_(D)=−22.8° (c=0.0029 in water), ES-MS 276 (M−1).

Preparation of 3-{[(1R)-1-(4-chlorophenyl)ethyl]amino}-1-propanesulfonicacid (Compound NV)

To a solution of (1R)-(−)-1-(4-chlorophenyl)ethylamine (5.07 g, 32.6mmol) in pinacolone (20 mL) and toluene (20 mL) was added 1,3-propanesultone (3.79 g, 31.0 mmol). The solution was stirred at reflux for 4hours. The reaction mixture was cooled to room temperature. The solidwas collected by filtration and was washed with acetone (2×25 mL). Thesolid was suspended in EtOH (60 mL). The suspension was stirred atreflux for 1 hour. The mixture was cooled to room temperature, the solidwas collected by filtration, washed with acetone (2×25 mL) and dried ina vacuum oven at 50° C., affording the title compound, 6.84 g (79%). ¹HNMR (D₂O, 500 MHz) δ ppm 7.39 (dd, 2H, J=2.4 Hz, 6.8 Hz), 7.32 (dd, 2H,J=2.4 Hz, 6.3 Hz), 4.31 (q, 1H, J=6.8 Hz), 3.00 (m, 1H), 2.84 (m, 1H),2.79 (t, 2H, J=7.3 Hz), 1.94 (m, 2H), 1.53 (d, 3H, J=6.8 Hz). ¹³C (D₂O,125 MHz) δ ppm 135.16, 134.41, 129.57, 129.36, 57.93, 48.02, 44.44,21.49, 18.11. [α]_(D)=+20.3° (c=0.0018 in water), ES-MS 276 (M−1).

Preparation of3-({1-[hydroxy(4-methylphenyl)methyl]cyclohexyl}amino)-1-propanesulfonicacid (Compound NW)

To a cooled solution of sodium methoxide (0.5 M in MeOH, 80 mL, 40mmol), nitrocyclohexane (5.0 g, 38.7 mmol) was added via syringe over a10 minute period. The reaction mixture was stirred at room temperaturefor 30 minutes and recooled before p-tolualdehyde (4.6 mL, 38.7 mmol)was added. The reaction mixture was stirred at room temperatureovernight. The mixture was neutralized with Amberlite IR-120 (stronglyacidic). The resin was removed by filtration and washed with MeOH (2×20mL). The filtrate was evaporated. The resulting oil was purified byflash chromatography: 98% Hexanes/EtOAc to 95% Hexanes/EtOAc, affordingthe desired nitro compound (1.2 g).

To a solution of the nitro compound (1.26 g, 5.0 mmol)) in MeOH wasadded 6M HCl (5 mL). After cooling to 5° C., zinc powder (1.63 g, 25.0mmol) was added. The suspension was stirred at room temperatureovernight. The mixture was filtered on a celite pad. The filter cake waswashed with MeOH (2×20 mL). The combined filtrates were evaporated underreduced pressure to afford the corresponding amine. The amine (1.03 g,67%) was used without further purification.

To a solution of amine (1.03 g, 4.7 mmol) in Pinacolone (9 mL) andToluene (9 mL) was added 1,3-propane sultone (558 mg, 4.5 mmol). Thesolution was stirred at reflux overnight. The reaction mixture wascooled to room temperature. The solid was collected by filtration, waswashed with acetone (2×15 mL) and dried in vacuo, affording the titlecompound, 930 mg (62%). ¹H NMR (DMSO, 500 MHz) δ ppm 8.26 (s (broad),1H), 8.13 (s (broad), 1H), 7.24 (d, 2H, J=8.3 Hz), 7.16 (d, 2H, J=7.8Hz), 6.19 (s, 1H), 4.74 (s, 1H), 3.22 (m, 1H), 3.11 (m, 1H), 2.72 (m,1H), 2.64 (m, 1H), 2.30 (s, 3H), 2.07 (m, 2H), 1.86 (m, 2H), 1.56 (m,2H), 1.41 (m, 3H), 1.21 (m, 2H), 0.88 (m, 1H). ¹³C (DMSO, 125 MHz) δ ppm148.33, 137.67, 129.07, 128.82, 73.18, 64.57, 50.11, 41.45, 28:13,27.65, 25.23, 23.46, 22.70, 21.42, 19.85, 19.58. ES-MS 340 (M−1).

Preparation of 3-{[(1S)-1-(4-methylphenyl)ethyl]amino}propane-1-sulfonicacid (Compound NX)

To a solution of (1S)-(−)-1-(4-methylphenyl)ethylamine (5.00 g, 37.0mmol) in Pinacolone (24 mL) and Toluene (24 mL) was added 1,3-propanesultone (4.30 g, 35.2 mmol). The solution was stirred at reflux for 4hours. The reaction mixture was cooled to room temperature. The solidwas collected by filtration and was washed with acetone (2×25 mL). Thesolid was suspended in EtOH (60 mL). The suspension was stirred atreflux for 1 hour. The mixture was cooled to room temperature, the solidmaterial was collected by filtration, washed with acetone (2×25 mL) anddried in a vacuum oven at 50° C., affording the title compound, 7.72 g(85%). ¹H NMR (D₂O, 500 MHz) δ ppm 7.22 (m, 4H), 4.26 (q, 1H, J=6.8 Hz),2.97 (m, 1H), 2.80 (m, 3H), 2.22 (s, 3H), 1.92 (m, 2H), 1.53 (d, 3H,J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 140.38, 132.79, 130.09, 127.71,58.34, 48.06, 44.34, 21.50, 20.41, 18.31. [α]_(D)=−26.4° (c=0.0019 inwater), ES-MS 256 (M−1).

Preparation of 3-{[(1R)-1-(4-methylphenyl)ethyl]amino}propane-1-sulfonicacid (Compound NY)

To a solution of (1R)-(+)-1-(4-methylphenyl)ethylamine (5.30 g, 39.1mmol) in pinacolone (25 mL) and toluene (25 mL) was added 1,3-propanesultone (4.55 g, 37.3 mmol). The solution was stirred at reflux for 4hours. The reaction mixture was cooled to room temperature. The solidwas collected by filtration and was washed with acetone (2×25 mL). Thesolid was suspended in EtOH (60 mL). The suspension was stirred atreflux for 1 hour. The mixture was cooled to room temperature, the solidmaterial was collected by filtration, washed with acetone (2×25 mL) anddried in a vacuum oven at 50° C., affording the title compound, 7.65 g(80%). ¹H NMR (D₂O, 500 MHz) δ ppm 7.21 (m, 4H), 4.25 (q, 1H, J=6.8 Hz),2.96 (m, 1H), 2.79 (m, 3H), 2.22 (s, 3H), 1.92 (m, 2H), 1.52 (d, 3H,J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 140.40, 132.79, 130.10, 127.72,58.34, 48.05, 44.34, 21.49, 20.39, 18.30. [α]_(D)=+28.8° (c=0.0025 inwater), ES-MS 256 (M−1).

Preparation of 3-[(cyclopropylmethyl)amino]-1-propanesulfonic acid(Compound NZ)

To a solution of cyclopropanemethylamine (5.12 g, 72.0 mmol) inPinacolone (40 mL) and Toluene (40 mL) was added 1,3-propane sultone(8.36 g, 68.7 mmol). The solution was stirred at reflux for 4 hours. Theproduct formed a sticky paste in the bottom of the flask. The reactionmixture was cooled to room temperature. The supernatant was removed. Theresidue was dissolved in a minimum of MeOH with heating. The solutionwas poured in acetone (300 mL) to precipitate the product. The solidmaterial was collected by filtration and washed with acetone (2×20 mL).The solid was suspended in EtOH (40 mL). The suspension was stirred atreflux for 1 hour. The mixture was cooled to room temperature, the solidmaterial was collected by filtration, washed with acetone (2×15 mL) anddried in a vacuum oven at 50° C., affording the title compound, 3.48 g(27%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.04 (t, 2H, J=7.8 Hz), 2.81 (m, 4H),1.96 (m, 2H), 0.90 (m, 1H), 0.51 (m, 2H), 0.18 (m, 2H). ¹³C (D₂O, 125MHz) δ ppm 52.82, 48.21, 46.01, 21.75, 7.09, 3.83. ES-MS 192 (M−1).

Preparation of3-{[(1R)-1-(3-methoxyphenyl)ethyl]amino}propane-1-sulfonic acid(Compound OA)

To a solution of (1S)-(−)-1-(3-methoxyphenyl)ethylamine (5.00 g, 33.1mmol) in pinacolone (20 mL) and toluene (20 mL) was added 1,3-propanesultone (3.84 g, 31.5 mmol). The solution was stirred at reflux for 4hours. The product formed a sticky paste in the bottom of the flask. Thereaction mixture was cooled to room temperature. The supernatant wasremoved. The residue was dissolved in MeOH with heating. Acetone (3×50mL) was added to precipitate the product. The solid material wascollected by filtration and washed with acetone (2×20 mL). The solid wassuspended in EtOH (60 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×25 mL) and dried in avacuum oven at 50° C., affording the title compound, 3.49 g (41%). ¹HNMR (D₂O, 500 MHz) δ ppm 7.27 (t, 1H, J=8.0 Hz), 6.90 (m, 3H), 4.22 (q,1H, J=6.7 Hz), 3.68 (s, 3H), 2.93 (m, 1H), 2.77 (m, 3H), 1.90 (m, 2H),1.49 (d, 3H, J=6.7 Hz). ¹³C (D₂O, 125 MHz) δ ppm 159.41, 137.34, 130.81,120.10, 115.24, 113.27, 58.65, 55.72, 48.18, 44.62, 21.76, 18.65.[α]_(D)=−23.0° (c=0.0019 in water), ES-MS 272 (M−1).

Preparation of 3-{[(1S)-1-phenylpropyl]amino}-1-propanesulfonic acid(Compound OB)

To a solution of (S)-(−)-1-phenylpropylamine (10.0 g, 74.1 mmol) inpinacolone (40 mL) and toluene (40 mL) was added 1,3-propane sultone(8.60 g, 70.6 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×25 mL). The solid wassuspended in EtOH (80 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×25 mL) and dried in avacuum oven at 50° C., affording the title compound, 14.38 g (79%). ¹HNMR (D₂O, 500 MHz) δ ppm 7.32 (m, 5H), 4.00 (dd, 1H, J=4.4 Hz, 10.7 Hz),2.91 (m, 1H), 2.74 (m, 3H), 1.90 (m, 4H), 0.62 (t, 3H, J=7.3 Hz). ¹³C(D₂O, 125 MHz) δ ppm 133.85, 129.89, 129.56, 128.41, 64.46, 48.01,44.48, 25.77, 21.40, 9.46. [α]_(D)=−15.6° (c=0.00077 in water), ES-MS256 (M−1).

Preparation of 3-{[(1R)-(1-naphthyl)ethyl]amino}-1-propanesulfonic acid(Compound OD)

To a solution of (R)-(+)-1-(naphthyl)ethylamine (5.02 g, 29.3 mmol) inpinacolone (15 mL) and toluene (15 mL) was added 1,3-propane sultone(3.40 g, 27.9 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×25 mL). The solid wassuspended in EtOH (40 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×25 mL) and dried in avacuum oven at 50° C., affording the title compound, 6.46 g (79%). ¹HNMR (DMSO, 500 MHz) δ ppm 8.21 (d, 1H, J=8.3 Hz), 7.98 (m, 2H), 7.73 (d,1H, J=7.3 Hz), 7.59 (m, 3H), 5.27 (m, 1H), 3.15 (m, 1H), 2.95 (m, 1H),2.55 (m, 2H), 1.96 (m, 2H), 1.59 (d, 3H, J=6.8 Hz). ¹³C (DMSO, 125 MHz)δ ppm 134.86, 134.05, 130.84, 129.67, 129.57, 127.62, 126.92, 126.25,124.34, 123.35, 52.55, 49.96, 46.21, 22.64, 20.38. [α]_(D)=−45.5°(c=0.0010 in water), ES-MS 292 (M−1).

Preparation of 3-{[(1S)-(1-naphthyl)ethyl]amino}-1-propanesulfonic acid(Compound OE)

To a solution of (S)-(−)-1-(naphthyl)ethylamine (5.00 g, 29.2 mmol) inpinacolone (15 mL) and toluene (15 mL) was added 1,3-propane sultone(3.39 g, 27.8 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×25 mL). The solid wassuspended in EtOH (40 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×25 mL) and dried in avacuum oven at 50° C., affording the title compound, 6.34 g (78%). ¹HNMR (DMSO, 500 MHz) δ ppm 8.21 (d, 1H, J=8.3 Hz), 7.97 (m, 2H), 7.72 (d,1H, J=7.3 Hz), 7.59 (m, 3H), 5.27 (m, 1H), 3.15 (m, 1H), 2.95 (m, 1H),2.61 (m, 2H), 1.96 (m, 2H), 1.59 (d, 3H, J=6.8 Hz). ¹³C (DMSO, 125 MHz)δ ppm 134.85, 134.06, 130.86, 129.68, 129.58, 127.64, 126.92, 126.26,124.35, 123.36, 52.55, 49.92, 46.20, 22.63, 20.36. [α]_(D)=+47.3°(c=0.00047 in water), ES-MS 292 (M−1).

Preparation of 3-{[4-methoxy-1,1-dimethylbutyl]amino}-1-propanesulfonicacid (Compound OF)

To a stirred solution of the alcohol (500 mg, 3.40 mmol) in DMF (6 mL)was added iodomethane (423 μL, 6.80 mmol) followed by NaH (163 mg, 6.80mmol). The reaction mixture was stirred for 15 hours then diluted withHCl (1M) and with EtOAc. The organic layer was washed with HCl (1M) thenconcentrated under high vacuum. The crude was purified by column usingHex:EtOAc 80:20 to obtain 450 mg of the desired product (82% yield).

To a stirred solution of the nitro (400 mg, 2.45 mmol) in methanol (5mL) was added a small spatula of Raney-Ni in water. The suspension washydrogenated under atmospheric pressure of hydrogen for 3 hours (TLCindicates complete consumption of the starting material) then filteredon prewashed celite and concentrated under reduced pressure. The crudeamine was used as such in the next step.

To the crude amine (300 mg, 2.29 mmol) in solution in THF (5 mL) wasadded 1,3-sultone (300 mg, 2.52 mmol) and the mixture was heated atreflux for 15 hours. The suspension was cooled down and filtered. Thesolid was dried under high vacuum to afford 400 mg of the correspondinghomotaurin as a white solid (69% yield). ¹H NMR (500 MHz, DMSO-d6) 3 ppm1.21 (s, 6H), 1.56 (m, 4H), 1.95 (m, 2H), 2.65 (m, 2H), 3.00 (m, 2H),3.23 (s, 3H), 3.32 (m, 2H), 8.53 (bs, 2H). ¹³C NMR (125 MHz, DMSO-d₆) δppm 22.93, 23.43, 23.85, 35.23, 41.16, 50.10, 58.52, 58.75, 72.28. ES-MS328 (M−1).

Preparation of 3-[(1,1-dimethyl-3-oxobutyl)amino]-1-propanesulfonic acid(Compound OG)

A mixture of mesityl oxide (4 g, 40 mmol) and aq. NH₃ was stirred for 15hours then diluted with EtOAc. Nitrogen was blown through the solutionto remove the excess of ammonia. Water was added and the two phases wereseparated. The aqueous phase was extracted with CH₂Cl₂ and the twophases were combined, dried (Na₂SO₄) and concentrated under rotavap andpump vacuum.

The crude amine was dissolved in THF (20 mL) to which was added1,3-propane sultone (2.2 g, 13.13 mmol). The reaction mixture wasstirred at reflux for 6 hours then cooled to room temperature. The solidwas collected by filtration and was washed with THF. The solid wassuspended in EtOH (10 mL) and stirred at reflux for 1 hour. Thesuspension was then cooled to room temperature. The solid was collectedby filtration, washed with ethanol and dried under high vacuum to affordthe title compound, 1 g (10%, the low yield is due to partial solubilityof the final product in EtOH and Et₂O). ¹H NMR (500 MHz, D₂O) 1.27 (s,6H), 1.99 (m, 2H), 2.12 (s, 3H), 2.88 (m, 2H), 2.92 (s, 2H), 3.03 (m,2H), 4.65 (s, 2H). ¹³NMR (125 MHz, D₂O) δ 21.99, 22.58, 23.61, 30.72,40.30, 47.33, 47.98, 58.02, 110.00. ES-MS 236 (M−1).

Preparation of3-{[4-(benzyloxy)-1,1-dimethylbutyl]amino}-1-propanesulfonic acid(Compound OH)

To a stirred solution of the alcohol (500 mg, 3.40 mmol) in DMF (5 mL)was added benzyl bromide (456 uL, 3.74 mmol) followed by NaH (106 mg,4.42 mmol). The reaction mixture was stirred for 15 hours then dilutedwith HCl (1M) and EtOAc. The organic layer was washed with HCl (1M) thenconcentrated under high vacuum. The crude product was purified by columnusing Hex:EtOAc 90:10 to obtain 605 mg of the desired product (74%yield).

To a stirred solution of the nitro (237 mg, 1.2 mmol) in ethanol (8 mL)was added HCl (6N) (2 mL) followed by Zn-dust. The suspension wasstirred for 10 minutes, then filtered and concentrated. The crudereaction mixture was diluted with EtOAc and neutralized with saturatedK₂CO₃. The organic layer was washed with water, dried over Na₂SO₄ andconcentrated. The crude amine was used as such in the next step.

To the crude amine (500 mg, 2.41 mmol) in solution in THF (5 mL) wasadded 1,3-sultone (234 mg, 2.65 mmol) and the mixture was heated atreflux of THF for 15 hours. The suspension was cooled down and filtered.The solid was dried to afford 300 mg of the homotaurine as a white solid(38% yield). ¹H NMR (500 MHz, DMSO-d6) δ 1.21 (s, 6H), 1.60 (m, 4H),1.95 (m, 2H), 2.65 (m, 2H), 3.00 (m, 2H), 3.42 (m, 2H), 4.46 (s, 2H),7.25-7.38 (m, 5H), 8.48 (bs, 2H). ¹³C NMR (125 MHz, DMSO-d₆) δ 22.93,23.43, 24.04, 35.29, 41.18, 50.08, 58.79, 70.07, 72.56, 128.09, 128.19,128.96, 139.23. ES-MS 328 (M−1).

Preparation of 3-piperidinylmethanesulfonic acid (Compound OI)

A solution of 3-hydroxymethylpiperidine (15 g, 129 mmol) in anhydrousCHCl₃ (120 mL) was saturated with HCl(g) and then treated dropwise atreflux with SOCl₂ (24 mL). The resulting mixture was refluxed for 1 hourand concentrated to yield a white solid, which was collected byfiltration and washed with Et₂O. It was then dissolved in EtOH and thenrecrystallized in EtOH/Et₂O to obtain 22 g of the desired chloride (96%yield).

A solution of the chloride (21.5 g, 121 mmol) in water (30 mL) was addeddropwise to a refluxed solution of N₂SO₃ (30.41 g, 242 mmol) in water(120 mL). After the end of the addition, the reaction was stirred atreflux for 60 minutes then cooled down and concentrated under reducedpressure. 75 mL of HCl (cone) were added to dissolve the aminosulfonicacid and precipitate the inorganic salts which were removed byfiltration. The filtrate was concentrated, then ethanol was added tocause amino sulfonic acid to appear as white solid which was collectedby filtration. It was washed with EtOH and Et₂O, then dried under highvacuum to obtain 18 g of a white solid (88% yield). ¹H NMR (500 MHz,D₂O) δ 1.25 (ddd, J=12.0, 9.0 and 3.0 Hz, 1H), 1.55-1.65 (m, 1H), 1.82(m, 1H), 1.90 (m, 1H), 2.10-2.20 (m, 1H), 2.68 (m, 1H), 2.72-2.86 (m,3H), 3.25 (dd, J=12.0 and 3.0 Hz, 1H), 3.50 (dd, J=12.0 & 3.0 Hz, 1H),4.69 (s, 2H). ¹³NMR (125 MHz, D₂O) δ 21.78, 28.08, 30.67, 44.05, 47.75,54.07. ES-MS 178 (M−1).

Preparation of 3-[3-(hydroxymethyl)piperidin-1-yl]-1-propanesulfonicacid (Compound OJ)

To a stirred solution of the 3-piperidinemethanol (1.15 g, 10 mmol) inTHF (20 mL) was added 1,3-propane sultone (1.2 g, 10 mmol). The reactionmixture was stirred at reflux for 15 hours then cooled to roomtemperature. The solid was collected by filtration and was washed withTHF. The solid was suspended in EtOH (20 mL) and stirred at reflux for 1hour. The suspension was then cooled to room temperature. The solid wascollected by filtration, washed with ethanol and dried under high vacuumto afford the title compound, 2.12 g (90%). ¹H NMR (500 MHz, DMSO-d₆) δ1.12 (m, 1H), 1.50-1.85 (m, 5H), 1.55-1.75 (m, 2H), 1.80-1.95 (m, 2H),1.992-2.10 (m, 2H), 2.60 (m, 1H), 2.70-2.80 (m, 1H), 2.85 (t, J=9.0 Hz,2H), 3.15 (t, J=9.0 Hz, 2H), 3.35-3.50 (m, 2H), 4.65 (s, 1H). ¹³NMR (125MHz, DMSO-d₆) δ 19.58, 22.43, 24.34, 36.84, 48.01, 53.18, 55.10, 56.06,63.37. ES-MS 236 (M−1).

Preparation of 3-[2-(2-hydroxyethyl)piperidin-1-yl]-1-propanesulfonicacid (Compound OK)

To a stirred solution of the 2-piperidinethanol (1.3 g, 10 mmol) in THF(20 mL) was added 1,3-propane sultone (1.2 g, 10 mmol). The reactionmixture was stirred at reflux for 15 hours then cooled to roomtemperature. The solid was collected by filtration and was washed withTHF. The solid was suspended in EtOH (20 mL) and stirred at reflux for 1hour. The suspension was then cooled to room temperature. The solid wascollected by filtration, washed with ethanol and dried under high vacuumto afford the title compound, 2.10 g (84%). ¹H NMR (500 MHz, DMSO-d₆) δ1.42 (m, 2H), 1.50-1.85 (m, 5H), 1.90-2.10 (m, 3H), 2.95 (m, 1H),3.10-3.22 (m, 3H), 3.30-3.70 (m, 3H), 4.63 (s, 1H). ¹³NMR (125 MHz,DMSO-d₆) δ 19.03, 20.80, 22.37, 27.66, 32.20, 49.35, 51.49, 52.38,57.76, 61.16. ES-MS 250 (M−1).

Preparation of (S)-3-[1-(4-bromophenyl)ethylamino]-1-propanesulfonicacid (Compound OL)

A solution of 1,3-propane sultone (1M, 5 mL) in toluene was added to asolution of (S)-(−)-1-(4-bromophenyl)ethylamine (1 g, 5.00 mmol) in MTBK(5 mL). The mixture was heated to reflux for 4 hours then cooled to roomtemperature. The solid was collected by suction filtration, rinsed withacetone (2×5 mL). The solid was dried 18 hours at 60° C. in the vacuumoven. The title compound was obtained as a fine white solid (1.12 g,3.48 mmol, 70%). ¹H NMR (500 MHz, DMSO-d6) δ 1.50 (d, J=6.8 Hz, 3H),1.93 (qt, J=6.6 Hz, 2H), 2.61 (t, J=6.6 Hz, 2H), 2.79 (qt, J=6.3 Hz,2H), 3.01 (qt, J=6.3 Hz, 2H), 4.38 (q, J=6.7 Hz, 1H), 7.45 (d, J=8.8 Hz,1H), 7.67 (d, J=8.8 Hz, 2H), 9.06 (br s, 1H), 9.20 (br s, 1H); ¹³C NMR(125 MHz, DMSO-d6) 19.0, 21.9, 45.1, 49.1, 56.1, 122.3, 129.9, 131.9,136.6; ES-MS 320-322 (M−H); [α]_(D)=−34° (c=0.00401, 0.1 N NaOH).

Preparation of (S)-3-[1-(4-nitrophenyl)ethylamino]-1-propanesulfonicacid (Compound OM)

A solution of 1,3-propane sultone (1M, 5.40 mL) in toluene was added toa solution of (S)-(−)-1-(4-nitrophenyl)ethylamine (0.895 g, 5.39 mmol)in MTBK (5 mL). The mixture was heated to reflux for 4 hours then cooledto room temperature. The solid was collected by suction filtration,rinsed with acetone (2×5 mL). The solid was dried 18 hours at 60° C. inthe vacuum oven. The title compound was obtained as a fine white solid(0.73 g, 2.53 mmol, 47%). ¹H NMR (500 MHz, DMSO-d6) δ 1.55 (d, J=6.8 Hz,3H), 1.95 (qt, J=6.6 Hz, 2H), 2.63 (t, J=6.6 Hz, 2H), 2.84 (qt, J=6.3Hz, 2H), 3.07 (qt, J=6.2 Hz, 2H), 4.58 (q, J=6.4 Hz, 1H), 7.78 (d, J=8.8Hz, 1H), 8.33 (d, J=8.8 Hz, 2H), 9.23 (br s, 1H), 9.38 (br s, 1H); ¹³CNMR (125 MHz, DMSO-d6) 18.9, 21.9, 45.3, 49.1, 56.0, 124.1, 129.1,144.5, 147.8; ES-MS 287 (M−H); [α]_(D)=−37°, c=0.0043, 0.1 N NaOH

Preparation of 3-(1-Carbamoyl-cyclohexylamino)-1-propanesulfonic acid(Compound ON)

To a 250 mL 1 neck flask containing 30% NH₄OH (120 mL) was added NaCN(15.34 g, 0.31 mol) and NH₄Cl (19.75 g, 0.37 mol) with vigorousstirring. The corresponding ketone was added dropwise within 20 minutesat room temperature. The mixture was stirred for 3 days at roomtemperature followed by extraction with dichloromehtnae (50 mL). Theorganic layer was separated and dried over anhydrous sodium sulfate for2 hours. The sodium sulfate was removed by filtration, the solvent wasremoved under reduced pressure to yield the crude aminonitrile. Thedesired material was obtained as a light brown oil (90% crude yield). ¹HNMR (500 MHz, CD₃OD-d6) δ 1.23-1.28 (m, 1H), 1.43-1.49 (m, 2H),1.51-1.60 (m, 2H), 1.69-1.73 (m, 1H), 1.78-1.83 (m, 2H), 2.02 (br s,1H), 2.05 (br s, 1H); ¹³C NMR (125 MHz, CD₃OD-d6) 24.2, 25.9, 38.9,52.9, 124.9; FT-IR 2221 cm⁻² (CN).

To 10 g of concentrated sulfuric acid stirred in an ice cooled waterbath was added dropwise a solution of the aminonitrille (41 mmol) in 30mL CH₂Cl₂, maintaining the internal temperature at 15° C. Then, the bathwas removed and the mixture heated to 40° C. for 1 hour. The mixture wascooled in ac ice bath and poured onto 200 g of crushed ice. The mixturewas made pH 7-8 with 28% aqueous NH₃ and extracted with EtOAc (3×100mL). The extracts were collected, dried (MgSO₄), and evaporated todryness”. The crude solid was recrystallized in EtOAc/Hex. The desiredmaterial was obtained as a white foamy solid 0.89 g, 6.26 mmol, 43%. ¹HNMR (500 MHz, DMSO-d6) δ 1.12-1.23 (m, 1H), 1.27 (br s, 1H), 1.30 (br s,1H), 1.40-1.45 (m, 2H), 1.49-1.27 (m, 3H), 1.67-1.72 (m, 4H), 7.34 (brs, 1H), 6.82 (br s, 1H); ¹³C NMR (125 MHz, DMSO-d6) 21.0, 25.4, 34.7,56.4, 180.3.

A solution of 1,3-propane sultone (1M, 6.20 mL) in toluene was added toa solution of 1-aminocyclohexanecarboxamide (0.880 g, 6.19 mmol) in MTBK(5 mL). The mixture was heated to reflux for 4 hours then cooled to roomtemperature. The solid was collected by suction filtration, rinsed withacetone (2×5 mL). The solid was dried 18 hours at 60° C. in the vacuumoven (0.79 g). The solid was recrystallized in ethanol (5 mL) and water(5 mL). After drying, the title compound was obtained as a fine whitesolid (0.4477 g, 1.69 mmol, 27%). ¹H NMR (500 MHz, DMSO-d6) δ 1.42 (brs, 4H), 1.63 (br s, 2H), 1.67-1.72 (m, 2H), 1.99 (qt, J=6.6 Hz, 2H),2.06-2.10 (m, 2H), 2.63 (t, J=6.6 Hz, 2H), 2.93 (br s, 2H), 7.69 (s,1H), 7.80 (s, 1H), 8.77 (br s, 2H); ¹³C NMR (125 MHz, DMSO-d6) 20.7,22.1, 24.1, 29.9, 42.5, 49.0, 64.4, 170.5; ES-MS 263 (M−H).

Preparation of 3-{[(1R)-1-(2-naphthyl)ethyl]amino}-1-propanesulfonicacid (Compound OO)

To a solution of (R)-(+)-1-(2-naphthyl)ethylamine (5.00 g, 29.2 mmol) inpinacolone (20 mL) and toluene (15 mL) was added 1,3-propane sultone(3.39 g, 27.8 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×20 mL). The solid wassuspended in EtOH (40 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×20 mL) and dried in avacuum oven at 50° C., affording the title compound, 7.36 g (90%). ¹HNMR (DMSO, 500 MHz) δ ppm 9.15 (s (broad), 1H), 8.01 (m, 2H), 7.93 (m,2H), 7.62 (d, 1H, J=8.3 Hz), 7.56 (m, 2H), 4.52 (m, 1H), 3.03 (m, 1H),2.81 (m, 1H), 2.60 (m, 2H), 1.94 (m, 2H), 1.60 (d, 3H, J=6.8 Hz). ¹³C(DMSO, 125 MHz) δ ppm 135.30, 133.60, 133.34, 129.51, 128.67, 128.37,127.91, 127.45, 125.22, 57.67, 49.92, 45.91, 22.56, 19.87.[α]_(D)=+15.2° (c=0.00084 in water), ES-MS 292 (M−1).

Preparation of 3-{[(1SR)-1-(2-naphthyl)ethyl]amino}-1-propanesulfonicacid (Compound OP)

To a solution of (S)-(−)-1-(2-naphthyl)ethylamine (5.00 g, 29.2 mmol) inpinacolone (20 mL) and toluene (15 mL) was added 1,3-propane sultone(3.39 g, 27.8 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×20 mL). The solid wassuspended in EtOH (40 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×20 mL) and dried in avacuum oven at 50° C., affording the title compound, 7.62 g (93%). ¹HNMR (DMSO, 500 MHz) δ ppm 9.20 (s (broad), 1H), 8.01 (m, 2H), 7.95 (m,2H), 7.62 (d, 1H, J=8.3 Hz), 7.56 (m, 2H), 4.52 (m, 1H), 3.04 (m, 1H),2.81 (m, 1H), 2.61 (m, 2H), 1.93 (m, 2H), 1.60 (d, 3H, J=6.8 Hz). ¹³C(DMSO, 125 MHz) δ ppm 135.30, 133.60, 133.32, 129.51, 128.65, 128.37,127.90, 127.46, 125.20, 57.64, 49.95, 45.94, 22.55, 19.85.[α]_(D)=−17.3° (c=0.00052 in water), ES-MS 292 (M−1).

Preparation of (R)-(−)-3-(1-methylpropylamino)-1-propanesulfonic acid(Compound OQ)

A solution of 1,3-propane sultone (1.83 g, 15.0 mmol) in toluene (15 mL)was added to a solution of (R)-(−)-2-butylamine (1 g, 13.7 mmol) inacetone (10 mL). The mixture was heated to reflux for 24 hours. Themixture was cooled to room temperature, and the solid was collected bysuction filtration, rinsed with acetone (2×5 mL) and dried under vacuum(2.59 g). The solid was suspended in ethanol (17 mL) and the suspensionwas heated to reflux. Water (0.1 mL) was then added to afford a clearsolution. The mixture was slowly cooled to room temperature and thesolid was collected by suction filtration, rinsed with acetone (2×5 mL)and dried 2 hours at 60° C. in the vacuum oven. The title compound wasobtained as a fine white solid (2.39 g, 12.2 mmol, 89%). ¹H NMR (500MHz, D₂O) δ 0.82 (t, J=7.6 Hz, 3H), 1.14 (d, J=6.3 Hz, 3H), 1.39-1.46(m, 1H), 1.59-1.66 (m, 1H), 1.96 (q, J=7.7 Hz, 2H), 2.86 (t, J=7.3 Hz,2H), 3.00-3.12 (m, 3H); ¹³C NMR (125 MHz, D₂O) δ 8.8, 15.0, 21.6, 25.7,43.5, 48.1, 56.0; ES-MS 194 (M−H); [α]_(D)=−1.2 (c=0.0157, H₂O)

Preparation of 3-{[(1R)-1-phenylpropyl]amino}-1-propanesulfonic acid(Compound OR)

To a solution of (R)-(+)-1-phenylpropylamine (10.0 g, 74.1 mmol) inpinacolone (40 mL) and toluene (40 mL) was added 1,3-propane sultone(8.60 g, 70.4 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×25 mL). The solid wassuspended in EtOH (80 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×25 mL) and dried in avacuum oven at 50° C., affording the title compound, 13.11 g (72%). ¹HNMR (D₂O, 500 MHz) δ ppm 7.31 (m, 5H), 4.00 (dd, 1H, J=4.7 Hz, 10.5 Hz),2.89 (m, 1H), 2.74 (m, 3H), 1.89 (m, 4H), 0.62 (t, 3H, J=7.3 Hz). ¹³C(D₂O, 125 MHz) δ ppm 133.73, 129.80, 129.47, 128.32, 64.60, 48.21,44.70, 26.07, 21.72, 9.83. [α]_(D)=+15.4° (c=0.00081 in water), ES-MS256 (M−1).

Preparation of 3-(1-Carbamoyl-1-methylamino)-1-propanesulfonic acid(Compound OS)

To a 250 mL 1 neck flask containing 30% NH₄OH (120 mL) was added NaCN(15.34 g, 0.31 mol) and NH₄Cl (19.75 g, 0.37 mol) with vigorousstirring. The corresponding ketone was added dropwise within 20 minutesat room temperature. The mixture was stirred for 3 days at roomtemperature followed by extraction with dichloromehtnae (50 mL). Theorganic layer was separated and dried over anhydrous sodium sulfate for2 hours. The sodium sulfate was removed by filtration, the solvent wasremoved under reduced pressure to yield the crude aminonitrile. Thedesired material was obtained as an light brown oil (clear oil, 80%crude yield). Used as such. ¹H NMR (500 MHz, DMSO-d6) δ 1.35 (s, 6H),2.56 (s, 2H); ¹³C NMR (125 MHz, DMSO-d6) 28.94, 45.7, 125.9

To 10 g of concentrated sulfuric acid stirred in an ice cooled waterbath was added dropwise a solution of the aminonitrille (41 mmol) in 30mL CH₂Cl₂, maintaining the internal temperature at 15° C. Then the bathwas removed and the mixture heated to 40° C. for 1 hour. The mixture wascooled in an ice bath and poured onto 200 g of crushed ice. The mixturewas made pH 7-8 with 28% aqueous NH₃ and extracted with EtOAc (3×100mL). The extracts were collected, dried (MgSO₄), and evaporated todryness. The crude solid was recrystallized in EtOAc/Hex. The desiredmaterial was obtained as a white foamy solid 0.4462 g, 4.37 mmol, 4%. ¹HNMR (500 MHz, DMSO-d6) δ 1.15 (s, 6H), 1.82 (br s, 2H), 6.84 (br s, 1H),7.26 (br s, 1H); ¹³C NMR (125 MHz, DMSO-d6) 28.7, 54.1, 108.1

A solution of 1,3-propane sultone (1M, 4.30 mL) in toluene was added toa solution of 2-amino-2-methylpropaneamide (0.4350 g, 4.26 mmol) in MTBK(6 mL) and ethanol (0.5 mL). The mixture was heated to reflux for 4hours then cooled to room temperature. The solid was collected bysuction filtration, rinsed with acetone (2×5 mL). The solid was dried 18hours at 60° C. in the vacuum oven (0.56 g). The solid wasrecrystallizeed in ethanol (5 mL) and water (5 mL). After drying, thetitle compound was obtained as a fine white solid (0.3500 g, 1.56 mmol,37%). ¹H NMR (500 MHz, D₂O) δ 1.47 (s, 6H), 1.99 (qt, J=7.6 Hz, 2H),2.87 (t, J=7.3 Hz, 2H), 3.01 (t, J=7.8 Hz, 2H); ¹³C NMR (125 MHz, D₂O)21.7, 21.9, 42.0, 48.0, 62.5, 174.6; ES-MS 223 (M−H)

Preparation of 3-(1-Carbamoyl-1-cyclopentylamino)-1-propanesulfonic acid(Compound OT)

To a 250 mL 1 neck flask containing 30% NH₄OH (120 mL) was added NaCN(15.34 g, 0.31 mol) and NH₄Cl (19.75 g, 0.37 mol) with vigorousstirring. The corresponding ketone was added dropwise within 20 minutesat room temperature. The mixture was stirred for 3 days at roomtemperature followed by extraction with dichloromehtnae (50 mL). Theorganic layer was separated and dried over anhydrous sodium sulfate for2 hours. The sodium sulfate was removed by filtration, the solvent wasremoved under reduced pressure to yield the crude aminonitrile. Thedesired material was obtained as colorless oil after distillation underreduced pressure (14.03 g, 127 mmol, 51% yield). Used as such. ¹H NMR(500 MHz, DMSO-d6) δ 1.61-1.71 (m, 2H), 1.72-1.83 (m, 4H), 1.8-1.94 (m,2H), 2.42 (s, 2H); ¹³C NMR (125 MHz, DMSO-d6) 23.0, 40.0, 54.2, 125.7

To 10 g of concentrated sulfuric acid stirred in an ice cooled waterbath was added dropwise a solution of the aminonitrille (41 mmol) in 30mL CH₂Cl₂, maintaining the internal temperature at 15° C. Then the bathwas removed and the mixture heated to 40° C. for 1 hour. The mixture wascooled in ac ice bath and poured onto 200 g of crushed ice. The mixturewas made pH 7-8 with 28% aqueous NH₃ and extracted with EtOAc (3×100mL). The extracts were collected, dried (MgSO₄), and evaporated todryness. The crude solid was recrystallized in EtOAc/Hex. The desiredmaterial was obtained as a white solid 1.36 g, 10.6 mmol, 27%. ¹H NMR(500 MHz, DMSO-d6) δ 1.37-1.41 (m, 2H), 1.57-1.63 (m, 2H), 1.68-1.76 (m,2H), 1.78 (br s, 2H), 1.88-1.94 (m, 2H), 6.91 (br s, 1H), 7.40 (br s,1H); ¹³C NMR (125 MHz, DMSO-d6) 24.2, 39.3, 64.7, 180.0.

A solution of 1,3-propane sultone (1M, 6.30 mL) in toluene was added toa solution of 2-amino-2-methylpropaneamide (0.4350 g, 4.26 mmol) in MTBK(7 mL) and ethanol (0.5 mL). The mixture was heated to reflux for 4hours then cooled to room temperature. The solid was collected bysuction filtration, rinsed with acetone (2×5 mL). The solid was dried 18hours at 60° C. in the vacuum oven (0.74 g). The solid wasrecrystallized in ethanol (5 mL) and water (5 mL). After drying, thetitle compound was obtained as a fine white solid (0.39 g, 2.80 mmol,45%). ¹H NMR (500 MHz, D₂O) δ 1.72-1.76 (m, 4H), 1.91-2.02 (m, 4H),2.08-2.14-(m, 2H), 2.86 (t, J=7.3 Hz, 2H), 3.00 (t, J=7.6 Hz, 2H); ¹³CNMR (125 MHz, D₂O) 22.0, 24.6, 34.8, 43.3, 48.0, 72.4, 174.8; ES-MS 249(M−H)

Preparation of 3-(1-Carbamoyl-cycloheptylamino)-1-propanesulfonic acid(Compound OU)

To a 250 mL 1 neck flask containing 30% NH₄OH (120 mL) was added NaCN(15.34 g, 0.31 mol) and NH₄Cl (19.75 g, 0.37 mol) with vigorousstirring. The corresponding ketone was added dropwise within 20 minutesat room temperature. The mixture was stirred for 3 days at roomtemperature followed by extraction with dichloromehtnae (50 mL). Theorganic layer was separated and dried over anhydrous sodium sulfate for2 hours. The sodium sulfate was removed by filtration, the solvent wasremoved under reduced pressure to yield the crude aminonitrile. Thedesired material was obtained as light yellow oil (33.09 g, 239 mmol,96% crude yield). An attempt to purify it further by distillation underreduced pressure was not effective. The material obtained after thedistillation was less pure than the crude product by was used as such inthe nest step. ¹H NMR (500 MHz, DMSO-d6) δ 1.41-67 (m, 10H), 1.91-95 (m,2H), 2.47 (s, 2H); ¹³C NMR (125 MHz, DMSO-d6) 21.8, 27.4, 40.1, 53.8,125.8

To 10 g of concentrated sulfuric acid stirred in an ice cooled waterbath was added dropwise a solution of the aminonitrille (41 mmol) in 30mL CH₂Cl₂, maintaining the internal temperature at 15° C. Then the bathwas removed and the mixture heated to 40° C. for 1 hour. The mixture wascooled in ac ice bath and poured onto 200 g of crushed ice. The mixturewas made pH 7-8 with 28% aqueous NH₃ and extracted with EtOAc (3×100mL). The extracts were collected, dried (MgSO₄), and evaporated todryness. The crude solid was recrystallized in EtOAc/Hex. The desiredmaterial was obtained as a white solid 2.15 g, 13.8 mmol, 31%. ¹H NMR(500 MHz, DMSO-d6) δ 1.34-1.38 (m, 2H), 1.49 (br s, 8H), 1.74 (s, 2H),1.88-1.93 (m, 2H), 6.76 (br s, 1H), 7.27 (br s, 1H); ¹³C NMR (125 MHz,DMSO-d6) 22.4, 29.6, 39.1, 59.6, 181.1

A solution of 1,3-propane sultone (1M, 5.20 mL) in toluene was added toa solution of 2-amino-2-methylpropaneamide (0.4350 g, 4.26 mmol) in MTBK(5 mL) and ethanol (0.5 mL). The mixture was heated to reflux for 4hours then cooled to room temperature. The solid was collected bysuction filtration, rinsed with acetone (2×5 mL). The solid was dried 18hours at 60° C. in the vacuum oven (0.62 g). The solid wasrecrystallized in ethanol (5 mL) and water (5 mL). After drying, thetitle compound was obtained as a fine white solid (0.39 g, 1.40 mmol,27%). ¹H NMR (500 MHz, DMSO-d6) δ 1.53 (br s, 6H), 1.65-1.69 (m, 2H),1.83-1.88-(m, 2H), 1.97 (qt, J=6.7 Hz, 2H), 2.06-2.11 (m, 2H), 2.61 (t,J=6.6 Hz, 2H), 2.91 (br s, 2H), 7.57 (s, 1H0, 7.79 (s, 1H), 8.75 (br s,2H); ¹³C NMR (125 MHz, DMSO-d6) 21.8, 22.2, 29.1, 33.1, 43.0, 49.1,68.3, 172.0; ES-MS 277 (M−H)

Preparation of 3-(1,4-dimethyl-pentylamino)-1-propanesulfonic acid(Compound OV)

At reflux, a solution of 1,3-propane sultone (10.90 g, 89 mmol) intoluene (75 mL) was added dropwise over a 20 minute period to a solutionof to a solution of 2-amino-5-methylhexane (10.00 g, 88.6 mmol) inacetone (80 mL). The mixture was heated to reflux for 7 hours then leftat room temperature for the night. The solid was collected by suctionfiltration, rinsed with acetone (2×25 mL). The solid was dried 1 hour at60° C. in the vacuum oven (15.20 g). The solid was recrystallized inmethanol (90 mL) and water (5 mL). The mixture was left to cool to roomtemperature with stirring. The solid was collected by suctionfiltration, rinsed with methanol (2×15 mL). The solid was dried 18 hoursat 60° C. in the vacuum oven. The title compound was obtained as a finewhite solid (12.67 g, 53.38 mmol, 60%). ¹H NMR (500 MHz, D₂O) δ ppm 0.88(dd, J=6.6, 2.2 Hz, 6H), 1.22-1.27 (m, 2H), 1.29 (d, J=6.3 Hz, 3H),1.51-1.60 (m, 2H), 1.71-1.78 (m, 1H), 2.08-2.14 (m, 2H), 3.00 (t, J=7.3Hz, 2H), 3.15-3.24 (m, 2H), 3.25-3.31 (m, 1H); ¹³C NMR (125 MHz, D₂O) δppm 15.5, 21.5, 21.6, 21.9, 27.3, 30.4, 33.5, 43.4, 48.1, 55.1; ES-MS236 (M−H)

Preparation of 3-(1,5-dimethyl-hexylamino)-1-propanesulfonic acid(Compound OW)

At reflux, a solution of 1,3-propane sultone (9.80 g, 80 mmol) intoluene (70 mL) was added dropwise over a 30 minute period to a solutionof to a solution of 2-amino-6-methylheptane (10.00 g, 78 mmol) inacetone (75 mL). The mixture was heated to reflux for 7 hours then itwas left at room temperature for the night. The solid was collected bysuction filtration, rinsed with acetone (2×20 mL). The solid was dried 1hour at 60° C. in the vacuum oven (15.20 g). The solid wasrecrystallized in methanol (45 mL). The mixture was left to cool to roomtemperature without stirring. The lump was crushed and diluted withacetone. The solid was collected by suction filtration, rinsed withacetone (2×25 mL). The solid was dried 18 hours at 60° C. in the vacuumoven. The title compound was obtained as a white solid (14.10 g, 56.10mmol, 72%). ¹H NMR (500 MHz, D₂O) δ 0.86 (dd, J=6.6, 2.0 Hz, 6H), 1.20(q, J=7.3 Hz, 2H), 1.30 (d, J=6.8 Hz, 3H), 1.32-1.45 (m, 2H), 1.49-1.58(m, 2H), 1.68-1.75 (m, 1H), 2.07-2.16 (m, 2H), 3.01 (t, J=7.3 Hz, 2H),3.15-3.25 (m, 2H), 3.27-3.34 (m, 1H); ¹³C NMR (125 MHz, D₂O) 15.5, 21.6,21.8, 22.0, 22.4, 27.2, 32.7, 37.9, 43.4, 48.1, 54.9; ES-MS 250 (M−H)

Preparation of 3-(1-methyl-butylamino)-1-propanesulfonic acid (CompoundOX)

At reflux, a solution of 1,3-propane sultone (10.05 g, 115 mmol) intoluene (100 mL) was added dropwise over a 30 minutes period to asolution of to a solution of 2-aminopentane (10.00 g, 115 mmol) inacetone (100 mL). The mixture was heated to reflux for 24 hours then wascooled to 0° C. with an ice/water bath. The solid was collected bysuction filtration, rinsed with acetone (2×20 mL). The solid was dried 1hours at 60° C. in the vacuum oven (19.42 g). The solid wasrecrystallized in methanol (45 mL). The mixture was left to cool to roomtemperature without stirring. The lump was crushed and diluted withacetone. The solid was collected by suction filtration, rinsed withethanol (2×20 mL). The solid was dried 3 days at 60° C. in the vacuumoven. The title compound was obtained as a white solid (15.53 g, 74.20mmol, 65%). ¹H NMR (500 MHz, D₂O) δ 0.92 (t, J=7.3 Hz, 3H), 1.29 (d,J=6.3 Hz, 3H), 1.31-1.47 (m, 2H), 1.50-1.57 (m, 1H), 1.67-1.74 (m, 1H),2.06-2.16 (m, 2H), 3.01 (t, J=7.3 Hz, 2H), 3.15-3.26 (m, 2H), 3.28-3.34(m, 1H); ¹³C NMR (125 MHz, D₂O) 13.1, 15.4, 18.1, 21.6, 34.7, 43.4,48.1, 54.6; ES-MS 208 (M−H)

Preparation of (R)-(+)-3-(1-methyl-octylamino)-1-propanesulfonic acid(Compound OY)

At reflux, a solution of 1,3-propane sultone (7.46 g, 39.0 mmol) intoluene (30 mL) was added to a solution of to a solution of(R)-(−)-2-aminononane (5.49 g, 38.3 mmol) in acetone (30 mL). Themixture was heated under reflux for 8 hours then at room temperature forthe weekend. Ether (20 mL) was added then the solid was collected bysuction filtration, rinsed with acetone (2×10 mL). The solid was dried 1hours at 60° C. in the vacuum oven (7.97 g). The solid was suspended inethanol (40 mL) and heated to reflux for 90 minutes. The mixture wasthen cooled to 0° C. The solid was collected by suction filtration,rinsed with ethanol (2×10 mL). The solid was dried 18 hours at 60° C. inthe vacuum oven. The title compound was obtained as a fine white solid(7.82 g, 29.5 mmol, 77%). ¹H NMR (500 MHz, DMSO-d6) δ 0.87 (t, J=6.8 Hz,3H), 1.16 (d, J=6.8 Hz, 3H), 1.26-1.40 (m, 11H), 1.60-1.65 (m, 1H), 1.93(qt, J=6.6 Hz, 2H), 2.64 (t, J=6.6 Hz, 2H), 3.04-3.11 (m, 3H), 8.47 (brs, 2H); ¹³C NMR (125 MHz, DMSO-d6) δ 13.9, 15.7, 21.9, 22.1, 24.5, 28.5,28.7, 31.2, 32.5, 44.0, 49.2, 53.1; ES-MS 264 (M−H); [α]_(D)=1.34±0.14(c=0.008753 in 0.1N NaOH)

Preparation of 3-{[1-(3,5-dimethoxy)cyclohexyl]amino}-1-propanesulfonicacid (Compound OZ)

NaOMe (0.5M, 40 mL) was added to nitrocyclohexane (2.58 g, 20 mmol) andthe solution was stirred for 30 minutes then concentrated to afford awhite solid. To this solid was added 3,5-dimethoxybenzylpyrridinium(5.45 g, 10 mmol) and DMSO (20 mL). The mixture was heated at 100° C.for 15 hours then cooled to room temperature and diluted with HCl (1M)and EtOAc. After separation of the two phases, the organic layer waswashed twice with HCl (1M) then concentrated to obtain an oily crude,mixed with some solid. Methanol was added to precipitate the pyridiniumbyproduct which was filtered off, and the filtrate was concentratedunder high vacuum. The crude was submitted to hydrogenation withoutfurther purification.

To a stirred solution of the crude nitro in methanol (20 mL) was added aspatula of Raney-Ni in water. The suspension was hydrogenated underatmospheric pressure of hydrogen for 15 hours (TLC indicates completeconsumption of the starting material) then filtered on celite andconcentrated under reduced pressure. The crude was purified by columnusing CH₂Cl₂:MeOH 80:10 to afford 1.2 g of the corresponding amine.

To a stirred solution of the amine (800 mg, 3.20 mmol) in THF (10 mL)was added 1,3-propane sultone (390 mg, 3.20 mmol). The reaction mixturewas stirred at reflux for 15 hours then cooled to room temperature. Thesolid was collected by filtration and was washed with THF. The solid wassuspended in EtOH (10 mL) and stirred at reflux for 1 hour. Thesuspension was then cooled to room temperature. The solid was collectedby filtration, washed with ethanol and dried under high vacuum to affordthe title compound, 1.1 g (86%). ¹H NMR (500 MHz, DMSO-d₆) δ 1.31 (m,2H), 1.54-1.68 (m, 8H), 1.96 (m, 2H), 2.62 (m, 2H), 2.89 (s, 2H), 3.08(m, 2H), 6.32 (m, 2H), 6.42 (s, 1H), 8.39 (bs, 2H). ¹³NMR (125 MHz,DMSO-d₆) δ 20.92, 22.81, 25.07, 31.84, 41.04, 50.09, 55.82, 61.64,109.47, 161.01. ES-MS 370 (M−1).

Preparation of 3-{[1-(3,5-dimethoxy)cyclohexyl]amino}-1-propanesulfonicacid (Compound PA)

NaOMe (0.5M, 40 mL) was added to nitrocyclohexane (2.58 g, 20 mmol) andthe solution was stirred for 30 minutes then concentrated to afford awhite solid. To this solid was added 3,5-dimethoxybenzylpyrridinium(5.45 g, 10 mmol) and DMSO (20 mL). The mixture was heated at 100° C.for 15 hours then cooled to room temperature and diluted with HCl (1M)and EtOAc. After separation of the two phases, the organic layer waswashed twice with HCl (1 M) then concentrated to obtain an oily crude,mixed with some solid. Methanol was added to precipitate the pyridiniumbyproduct which was filtered off, and the filtrate was concentratedunder high vacuum. The crude was submitted to hydrogenation withoutfurther purification.

To a stirred solution of the crude nitro in methanol (20 mL) was added aspatula of Raney-Ni in water. The suspension was hydrogenated underatmospheric pressure of hydrogen for 15 hours and then was filtered oncelite and concentrated under reduced pressure. The crude was purifiedby column using CH₂Cl₂:MeOH 80:10 to afford 1.2 g of the correspondingamine.

To a stirred solution of the amine (800 mg, 3.20 mmol) in THF (10 mL)was added 1,3-propane sultone (390 mg, 3.20 mmol). The reaction mixturewas stirred at reflux for 15 hours then cooled to room temperature. Thesolid was collected by filtration and was washed with THF. The solid wassuspended in EtOH (10 mL) and stirred at reflux for 1 hour. Thesuspension was then cooled to room temperature. The solid was collectedby filtration, washed with ethanol and dried under high vacuum to affordthe title compound, 1.1 g (86%). ¹H NMR (500 MHz, DMSO-d₆) δ 1.31 (m,2H), 1.54-1.68 (m, 8H), 1.96 (m, 2H), 2.62 (m, 2H), 2.89 (s, 2H), 3.08(m, 2H), 6.32 (m, 2H), 6.42 (s, 1H), 8.39 (bs, 2H). ¹³NMR (125 MHz,DMSO-d₆) δ 20.92, 22.81, 25.07, 31.84, 41.04, 50.09, 55.82, 61.64,109.47, 161.01. ES-MS 370 (M−1).

Preparation of3-{[2-(3,5-dimethoxyphenyl)-1,1-dimethylethyl]amino}-1-propanesulfonicacid (Compound PB)

NaOMe (0.5M, 40 mL) was added to 2-nitropropane (1.78 g, 20 mmol) andthe solution was stirred for 30 minutes then concentrated to afford awhite solid. To this solid was added 3,5-dimethoxybenzylpyrridinium(5.45 g, 10 mmol) and DMSO (20 mL). The mixture was heated at 100° C.for 15 hours then cooled to room temperature and diluted with HCl (1M)and EtOAc. After separation of the two phases, the organic layer waswashed twice with HCl (1M) then concentrated to obtain an oily crude,mixed with some solid. Methanol was added to precipitate the pyridiniumbyproduct which was filtered off, and the filtrate was concentratedunder high vacuum. The crude was submitted to hydrogenation withoutfurther purification.

To a stirred solution of the crude nitro in methanol (20 mL) was added aspatula of Raney-Ni in water. The suspension was hydrogenated underatmospheric pressure of hydrogen for 15 hours (TLC indicates completeconsumption of the starting material) then filtered on celite andconcentrated under reduced pressure. The crude was purified by columnusing CH₂Cl₂:MeOH 80:10 to afford 1.2 g (57%) of the correspondingamine.

To a stirred solution of the amine (1.1 g, 5.25 mmol) inpinacolone/toluene (8 mL/2 mL) was added 1,3-propane sultone (642 mg,5.25 mmol). The reaction mixture was stirred at reflux for 15 hours thencooled to room temperature. The solid was collected by filtration andwas washed with THF. The solid was suspended in EtOH (10 mL) and stirredat reflux for 1 hour. The suspension was then cooled to roomtemperature. The solid was collected by filtration, washed with ethanoland dried under high vacuum to afford the title compound, 1.4 g (80%).¹H NMR (500 MHz, DMSO-d₆) δ 1.19 (s, 6H), 1.98 (m, 2H), 2.67 (m, 2H),2.82 (s, 2H), 3.11 (m, 2H), 3.74 (s, 6H), 6.38 (s, 2H), 6.42 (s, 1H),8.60 (bs, 2H). ¹³NMR (125 MHz, DMSO-d₆) δ 23.07, 23.53, 41.37, 44.20,49.86, 55.83, 59.28, 99.49, 109.49, 137.83, 160.91. ES-MS 330 (M−1).

Preparation of3-{[2-(2,4-dichlorophenyl)-1,1-dimethylethyl]amino}-1-propanesulfonicacid (Compound PD)

NaOMe (0.5M, 40 mL) was added to 2-nitropropane (1.78 g, 20 mmol) andthe solution was stirred for 30 minutes then concentrated to afford awhite solid. To this solid was added 2,4-dichlorobenzylpyrridinium (5.5g, 10 mmol) and DMSO (20 mL). The mixture was heated at 100° C. for 15hours then cooled to room temperature and diluted with HCl (1M) andEtOAc. After separation of the two phases, the organic layer was washedtwice with HCl (1M) then concentrated to obtain an oily crude, mixedwith some solid. Methanol was added to precipitate the pyridiniumbyproduct which was filtered off, and the filtrate was concentratedunder high vacuum. The crude was submitted to hydrogenation withoutfurther purification.

To a stirred solution of the crude nitro in methanol (20 mL) was added aspatula of Raney-Ni in water. The suspension was hydrogenated underatmospheric pressure of hydrogen for 15 hours (TLC indicates completeconsumption of the starting material) then filtered on celite andconcentrated under reduced pressure. The crude was purified by columnusing CH₂Cl₂:MeOH 80:10 to afford 980 mg (45%) of the correspondingamine.

To a stirred solution of the amine (960 mg, 4.40 mmol) inpinacolone/toluene (8 mL/2 mL) was added 1,3-propane sultone (537 mg,4.40 mmol). The reaction mixture was stirred at reflux for 15 hours thencooled to room temperature. The solid was collected by filtration andwas washed with THF. The solid was suspended in EtOH (10 mL) and stirredat reflux for 1 hour. The suspension was then cooled to roomtemperature. The solid was collected by filtration, washed with ethanoland dried under high vacuum to afford the title compound, 1.4 g (80%).¹H NMR (500 MHz, DMSO-d₆) δ1.20 (s, 6H), 2.00 (m, 2H), 2.70 (m, 2H),3.10 (s, 2H), 3.20 (m, 2H), 3.7.40 (s, 2H), 7.65 (s, 1H). ¹³NMR (125MHz, DMSO-d₆) δ 23.08, 23.18, 41.37, 49.83, 60.13, 128.04, 129.85,132.88, 133.49, 135.14, 135.90. ES-MS 340 & 338 (M−1).

Preparation of3-{[(2-(2,4-dichlorophenyl)-1,1-dimethylethyl]amino}-1-propanesulfonicacid (Compound PE)

NaOMe (0.5M, 40 mL) was added to 2-nitropropane (1.78 g, 20 mmol) andthe solution was stirred for 30 minutes then concentrated to afford awhite solid. To this solid was added 2,4-dichlorobenzylpyrridinium (5.5g, 10 mmol) and DMSO (20 mL). The mixture was heated at 100° C. for 15hours then cooled to room temperature and diluted with HCl (1M) andEtOAc. After separation of the two phases, the organic layer was washedtwice with HCl (1M) then concentrated to obtain an oily crude, mixedwith some solid. Methanol was added to precipitate the pyridiniumbyproduct which was filtered off, and the filtrate was concentratedunder high vacuum. The crude was submitted to hydrogenation withoutfurther purification.

To a stirred solution of the crude nitro in methanol (20 mL) was added aspatula of Raney-Ni in water. The suspension was hydrogenated underatmospheric pressure of hydrogen for 15 hours (TLC indicates completeconsumption of the starting material) then filtered on celite andconcentrated under reduced pressure. The crude was purified by columnusing CH₂Cl₂:MeOH 80:10 to afford 980 mg (45%) of the correspondingamine.

To a stirred solution of the amine (960 mg, 4.40 mmol) inpinacolone/toluene (8 mL/2 mL) was added 1,3-propane sultone (537 mg,4.40 mmol). The reaction mixture was stirred at reflux for 15 hours thencooled to room temperature. The solid was collected by filtration andwas washed with THF. The solid was suspended in EtOH (10 mL) and stirredat reflux for 1 hour. The suspension was then cooled to roomtemperature. The solid was collected by filtration, washed with ethanoland dried under high vacuum to afford the title compound, 1.4 g (80%).¹H NMR (500 MHz, DMSO-d₆) □ 1.20 (s, 6H), 2.00 (m, 2H), 2.70 (m, 2H),3.10 (s, 2H), 3.20 (m, 2H), 3.7.40 (s, 2H), 7.65 (s, 1H). ¹³NMR (125MHz, DMSO-d₆) □ 23.08, 23.18, 41.37, 49.83, 60.13, 128.04, 129.85,132.88, 133.49, 135.14, 135.90. ES-MS 340 & 338 (M−1).

Preparation of3-{[1,1-dimethyl-2-(4-propoxyphenyl)ethyl]amino}-1-propanesulfonic acid(Compound PF)

To a stirred solution of the phenol (195 mg, 1 mmol) in DMF (10 mL) wasadded NaH (48 mg, 2 mmol) followed by AllBr (170 μL, 2 mmol). Thesuspension was heated at reflux for 15 hours then diluted with HCl (1 M)and with EtOAc. The organic layer was washed with HCl(1 M) thenconcentrated under high vacuum to afford 200 mg of the desired product(97% yield).

To a stirred solution of the crude nitro in methanol (5 mL) was added asmall spatula of Raney-Ni in water. The suspension was hydrogenatedunder atmospheric pressure of hydrogen for 15 hours (TLC indicatescomplete consumption of the starting material) then filtered onpre-washed celite and concentrated under reduced pressure. The crudeamine was used as such in the next step.

To the crude amine (207 mg, 1 mmol) in solution in pinacolone (3 mL) wasadded 1,3-propane sultone (122 mg, 1 mmol) and the mixture was heated atreflux for 12 hours. The suspension of was cooled down and filtered. Thesolid was dried to afford 270 mg of the homotaurin as a white solid (79%yield). ¹H NMR (500 MHz, DMSO-d6) δ 0.97 (t, J=7.5 Hz, 3H), 1.14 (s,6H), 1.70 (m, 2H), 1.99 (m, 2H), 2.67 (m, 2H), 2.81 (s, 2H), 3.12 (m,2H), 3.90 (t, J=6.4 Hz, 2H), 6.88 (d, J=7.5 Hz, 2H), 7.12 (d, J=7.5 Hz,2H), 8.59 (bs, 2H). ¹³NMR (125 MHz, DMSO-d₆) δ 11.13, 22.75, 23.11,41.24, 43.06, 49.82, 59.44, 69.50, 114.86, 127.38, 132.37, 158.43. ES-MS328 (M−1).

Preparation of 2-piperidinylethanesulfonic acid (Compound PG)

A solution of 2-piperidineethanol (containing 10% of impurety) (6 g,46.44 mmol) in anhydrous CHCl₃ was saturated with HCl(g) and thentreated drop wise at reflux with SOCl₂. The resulting mixture wasrefluxed for 1 hour and concentrated to yield a brown solid. The solidwas dissolved in EtOH and then recrystallized in EtOH/Et₂O to obtain 6.4g of the desired chloride (76% yield).

A solution of the chloride (3.7 g, 20 mmol) in water (4 mL) was addeddrop wise to a refluxed solution of N₂SO₃ (5.04 g, 40 mmol) in water (18mL). After the end of the addition, the reaction was stirred at refluxfor 40 minutes then cooled down and concentrated under reduced pressure.12 mL of HCl(conc) were added to dissolve the aminosulfonic acid andprecipitate the inorganic salts which were removed by filtration. Thefiltrate was concentrated then ethanol was added to cause amino sulfonicacid to appear as white solid which was collected by filtration. It waswashed with EtOH and Et₂O, then dried under high vacuum to obtain 2.84 gof a white solid (73% yield). ¹H NMR (500 MHz, D₂O) δ 1.33-1.53 (m, 3H),1.75 (m, 2H), 1.82-1.92 (m, 2H), 1.93-2.00 (m, 1H), 2.80-2.92 (m, 3H),3.13 (m, 1H), 3.30 (m, 1H), 4.65 (s, 2H). ¹³NMR (125 MHz, D₂O) δ 21.59,21.94, 27.97, 28.60, 45.02, 46.79, 55.79. ES-MS 178 (M−1).

Preparation of 3-(1,1-Dimethyl-prop-2-ynylamino)-1-propanesulfonic acid(Compound PH)

The 1,3-propane sultone (1.22 g, 10 mmol) was added to a solution of1,1,-dimethylpropargylamine (0.8300 g, 10.00 mmol) in MeCN (15 mL). Themixture was heated to 75° C. for 4.5 hours then cooled to roomtemperature. The solid was collected by suction filtration, rinsed withacetone (2×5 mL). The solid was dried 18 hours at 60° C. in the vacuumoven. The title compound was obtained as a fine white solid (1.46 g,7.11 mmol, 71%). ¹H NMR (500 MHz, D₂O) δ 1.65 (s, 6H), 2.13 (qt, J=7.7Hz, 2H), 3.03 (t, J=7.6 Hz, 2H), 3.12 (s, 1H), 3.37 (t, J=7.8 Hz, 2H);¹³C NMR (125 MHz, D₂O) δ 21.8, 25.6, 41.7, 48.0, 54.1, 76.8, 50.5; ES-MS204 (M−H).

Preparation of (R)-(+)-3-[1-(4-bromophenyl)ethylamino]-1-propanesulfonicacid (Compound PI)

The 1,3-propane sultone (0.6110 g, 5 mmol) was added to a solution of(R)-(+)-1-(4-bromophenyl)ethylamine (1 g, 5.00 mmol) in MeCN (10 mL).The mixture was heated to 75° C. for 4.5 hours then cooled to roomtemperature. The solid was collected by suction filtration, rinsed withacetone (2×5 mL). The solid was dried 18 hours at 60° C. in the vacuumoven. The title compound was obtained as a fine white solid (1.63 g,5.06 mmol, 100%). ¹H NMR (500 MHz, DMSO-d6) δ 1.50 (d, J=6.8 Hz, 3H),1.93 (qt, J=6.6 Hz, 2H), 2.61 (t, J=6.6 Hz, 2H), 2.79 (qt, J=6.3 Hz,2H), 3.01 (qt, J=6.3 Hz, 2H), 4.38 (q, J=6.7 Hz, 1H), 7.45 (d, J=8.8 Hz,1H), 7.67 (d, J=8.8 Hz, 2H), 9.05 (br s, 1H), 9.21 (br s, 1H); ¹³C NMR(125 MHz, DMSO-d6) 19.0, 21.8, 45.1, 49.2, 56.1, 122.3, 129.9, 131.9,136.6; ES-MS 320-322 (M−H); [α]_(D)=+34.2±0.2° (c=0.003345 in 0.1 NNaOH)

Preparation of (S)-(+)-3-(1-methylpropylamino)-1-propanesulfonic acid(Compound PJ)

The 1,3-propane sultone (1.67 g, 13.7 mmol) was added to a solution of(S)-(+)-2-butylamine (1 g, 13.7 mmol) in a mixture of acetone (7 mL) andtoluene (7 mL). The mixture was heated to reflux for 6 hours. Themixture was cooled to room temperature, and the solid was collected bysuction filtration, rinsed with acetone (2×5 mL) and dried under vacuum.The title compound was obtained as a fine white solid (1.90 g, 9.73mmol, 71%). ¹H NMR (500 MHz, D₂O) δ 0.96 (t, J=7.6 Hz, 3H), 1.29 (d,J=6.3 Hz, 3H), 1.54-1.60 (m, 1H), 1.76-1.81 (m, 1H), 2.12 (q, J=7.7 Hz,2H), 3.01 (t, J=7.3 Hz, 2H), 3.15-3.27 (m, 3H); ¹³C NMR (125 MHz, D₂O) δ8.8, 14.9, 21.6, 25.7, 43.5, 48.1, 56.0; ES-MS 194 (M−H);[α]_(D)=1.10±0.04° (c=0.01364 in water)

Preparation of(−)-3-[(1R,2S,5R)-2-Isopropyl-5-methyl-cyclohexylamino]-1-propanesulfonicacid (Compound PK)

The 1,3-propane sultone (0.3940 g, 3.22 mmol) was added to a solution ofL-menthylamine (0.500 g, 3.22 mmol) in a mixture of acetone (3 mL) andtoluene (4 mL). The mixture was heated to reflux for 6 hours. Themixture was cooled to room temperature, and the solid was collected bysuction filtration, rinsed with acetone (2×5 mL) and dried 2 hours inthe vacuum oven at 60° C. (0.68 g). The solid was recrystallized inethanol (7 mL). The mixture was cooled to room temperature, and thesolid was collected by suction filtration, rinsed with acetone (2×5 mL)and dried 18 hours in the vacuum oven at 60° C. The title compound wasobtained as a fine white solid (0.3200 g, 1.15 mmol, 36%). ¹H NMR (500MHz, DMSO-d6) δ 0.75 (d, J=6.8 Hz, 3H), 0.78-1.07 (m, 3H), 0.91 (d,J=6.3 Hz, 6H), 1.31-1.35 (m, 1H0, 1.40-1.42 (m, 1H), 1.95-2.03 (m, 4H),2.63-2.70 (m, 2H), 2.95-3.05 (m, 2H), 3.10-3.20 (m, 1H), 8.40 (br s,1H), 8.65 (br s, 1H); ¹³C NMR (125 MHz, DMSO-d6) 8 15.4, 21.0, 21.8,22.0, 22.4, 24.5, 30.8, 33.3, 36.1, 43.4, 44.4, 49.6, 56.8; ES-MS 276(M−H); [α]_(D)=−46.2±0.3° (c=0.0568 in water).

Preparation of 3-{[(1S)-1-methylpentyl]amino}-1-propanesulfonic acid(Compound PL)

To a solution of (S)-2-aminohexane (5.10 g, 50.4 mmol) in acetone (20mL) and toluene (20 mL) was added 1,3-propane sultone (5.85 g, 48.0mmol). The solution was stirred at reflux for 4 hours. The reactionmixture was cooled to room temperature. The solid material was collectedby filtration and washed with acetone (2×20 mL). The solid was suspendedin EtOH (40 mL). The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature, the solid material was collectedby filtration, washed with acetone (2×20 mL) and dried in a vacuum ovenat 50° C., affording the title compound, 7.86 g (73%). ¹H NMR (D₂O, 500MHz) δ ppm 3.06 (m, 3H), 2.83 (t, 2H, J=7.3 Hz), 1.93 (quintet, 2H,J=7.4 Hz), 1.56 (m, 1H), 1.37 (m, 1H), 1.16 (m, 7H), 0.71 (m, 3H). ¹³C(D₂O, 125 MHz) δ ppm 54.96, 48.26, 43.57, 32.46, 26.97, 22.16, 21.91,15.79, 13.54. [α]_(D)=−6.3° (c=0.0051 in water), ES-MS 222 (M−1).

Preparation of 3-{[(1SR)-1-methylpentyl]amino}-1-propanesulfonic acid(Compound PM)

To a solution of (R)-2-aminohexane (5.12 g, 50.6 mmol) in acetone (20mL) and toluene (20 was added 1,3-propane sultone (5.87 g, 48.2 mmol).The solution was stirred at reflux for 4 hours. The reaction mixture wascooled to room temperature. The solid material was collected byfiltration and washed with acetone (2×20 mL). The solid was suspended inEtOH (40 mL). The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature, the solid material was collectedby filtration, washed with acetone (2×20 mL) and dried in a vacuum ovenat 50° C., affording the title compound, 7.66 g (71%). ¹H NMR (D₂O, 500MHz) δ ppm 3.01 (m, 3H), 2.86 (t, 2H, J=7.3 Hz), 1.97 (m, 2H), 1.59 (m,1H), 1.40 (m, 1H), 1.19 (m, 7H), 0.74 (t, 3H, J=6.8 Hz). ¹³C (D₂O, 125MHz) δ ppm 54.83, 48.08, 43.38, 32.22, 26.70, 21.87, 21.64, 15.50,13.21. [α]_(D)=+6.4° (c=0.0025 in water), ES-MS 222 (M−1).

Preparation of 3-{[(1S)-1,2-dimethylpropyl]amino}-1-propanesulfonic acid(Compound PN)

To a solution of (S)-(+)-3-methyl-2-butylamine (5.00 g, 57.4 mmol) inacetone (35 mL) and toluene (35 mL) was added 1,3-propane sultone (6.68g, 54.7 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×25 mL). The solid wassuspended in EtOH (40 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×25 mL) and dried in avacuum oven at 50° C., affording the title compound, 4.95 g (43%). ¹HNMR (D₂O, 500 MHz) δ ppm 3.07 (m, 3H), 2.87 (td, 2H, J=1.2 Hz, 7.3 Hz),1.95 (m, 3H), 1.07 (d, 3H, J=6.8 Hz), 0.83 (d, 3H, J=6.8 Hz), 0.78 (t,3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 59.76, 48.16, 44.15, 29.71,21.49, 18.42, 14.99, 10.66. [α]_(D)=+2.3° (c=0.0015 in water), ES-MS 208(M−1).

Preparation of 3-{[(1R)-1,2,2-trimethylpropyl]amino}-1-propanesulfonicacid (Compound PO)

To a solution of (R)-3,3-dimethyl-2-butylamine (10.0 g, 98.8 mmol) inacetone (40 mL) and toluene (40 mL) was added 1,3-propane sultone (11.5g, 94.1 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×25 mL). The solid wasrecrystallized in EtOH. After the crystals were filtered and washed withacetone (2×25 mL), the product was dried in a vacuum oven at 50° C.,affording the title compound, 11.94 g (57%). ¹H NMR (D₂O, 500 MHz) δ ppm3.20 (m, 1H), 3.05 (m, 1H), 2.95 (quartet, 1H, J=6.8 Hz), 2.89 (t, 2H,J=6.8 Hz), 2.02 (m, 2H), 1.13 (d, 3H, J=6.8 Hz), 0.86 (s, 9H). ¹³C (D₂O,125 MHz) δ ppm 64.03, 48.30, 45.42, 33.20, 25.11, 21.15, 11.15.[α]_(D)=−28.6° (c=0.0026 in water), ES-MS 222 (M−1).

Preparation of 3-{[(1R)-1,2-dimethylpropyl]amino}-1-propanesulfonic acid(Compound PP)

To a solution of (R)-(−)-3-methyl-2-butylamine (10.0 g, 115 mmol) inacetone (70 mL) and toluene (70 mL) was added 1,3-propane sultone (13.4g, 110 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×25 mL). The solid wassuspended in EtOH (40 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×25 mL) and dried in avacuum oven at 50° C., affording the title compound, 11.36 g (49%). ¹HNMR (D₂O, 500 MHz) δ ppm 3.07 (m, 3H), 2.86 (td, 2H, J=1.2 Hz, 7.3 Hz),1.95 (m, 3H), 1.07 (d, 3H, J=6.8 Hz), 0.83 (d, 3H, J=6.8 Hz), 0.78 (t,3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm 59.77, 48.17, 44.16, 29.72,21.50, 18.43, 15.00, 10.67. [α]_(D)=−2.9° (c=0.0026 in water), ES-MS 208(M−1).

Preparation of 3-(1-Methyl-3-phenylpropylamino)-1-propanesulfonic acid(Compound PQ)

A solution of 1,3-propane sultone (8.60 g, 70.40 mmol) in toluene (35mL) was added to a solution of 3-amino-1-phenylbutane (10.50 g, 70.36mmol) in acetone (35 mL). The mixture was heated to reflux for 4 hours.The mixture was cooled to room temperature, and the solid was collectedby suction filtration, rinsed with acetone (2×20 mL) and dried 1 hour inthe vacuum oven at 60° C. (14.02 g). The solid was suspended in ethanol(90 mL) and the mixture was heated 1 hours at reflux. The mixture wascooled to room temperature, and the solid was collected by suctionfiltration, rinsed with ethanol (2×15 mL) and dried 18 hours in thevacuum oven at 60° C. The title compound was obtained as a fine whitesolid (13.95 g, 51.40 mmol, 73%). ¹H NMR (500 MHz, DMSO-d6) δ 1.25 (d,J=6.8 Hz, 3H), 1.67-1.74 (m, 1H), 1.92-2.01 (m, 3H), 2.57-2.72 (m, 4H),3.04-3.18 (m, 3H), 7.19-7.25 (m, 3H), 7.29-7.32 (m, 2H), 8.56 (br s,2H); ¹³C NMR (125 MHz, DMSO-d6) δ 15.7, 22.0, 30.7, 34.3, 43.9, 49.1,52.9, 126.1, 128.3, 128.5, 140.8; ES-MS 270 (M−H)

Preparation of3-({1-[hydroxy(3-methoxyphenyl)methyl]cyclopentyl}amino)-1-propanesulfonicacid (Compound PR)

To a cooled solution of sodium methoxide (0.5 M in MeOH, 20 mL) wasadded via syringe over a 10 minutes period 2-nitrocyclopentane (3.00 g,26 mmol). The reaction mixture was stirred at room temperature for 30minutes and recooled before m-anisaldehyde (3.2 mL, 26 mmol) was added.The reaction mixture was stirred at room temperature overnight. Themixture was neutralized with Amberlite IR-120 (strongly acidic). Theresin was removed by filtration and washed with MeOH (2×15 mL). Thefiltrate was evaporated. The resulting oil was purified by flashchromatography: 100% Hexanes to 90% Hexanes/EtOAc, affording the desirednitro compound (1.6 g, 22%).

To a solution of the nitro compound (1.6 g, 5.6 mmol)) in MeOH (12 mL)was added 6M HCl (7 mL). After cooling to 5° C., zinc powder (1.85 g,28.2 mmol) was added. The suspension was stirred at 0-5° C. for 30minutes and at room temperature for 6 h. The mixture was filtered on acelite pad. The filter cake was washed with MeOH (2×15 mL). The combinedfiltrates were evaporated under reduced pressure. The residue wasdissolved in EtOAc (40 mL). The mixture was extracted with 5% NaOH (1×40mL). The aqueous phase was extracted with EtOAc (2×40 mL). The combinedorganic extracts were dried with Na₂SO₄, filtered, evaporated and driedin vacuo to afford the corresponding amine. The amine (0.920 g, 66%) wasused without further purification.

To a solution of amine (1.31 g, 6.3 mmol) in acetone (5 mL) and toluene(5 mL) was added 1,3-propane sultone (0.422 g, 3.4 mmol). The solutionwas stirred at reflux overnight. The reaction mixture was cooled to roomtemperature. The solid material was collected by filtration, was washedwith acetone (2×15 mL). The solid was suspended in EtOH (15 mL). Thesuspension was stirred at reflux for 1 hour. The mixture was cooled toroom temperature. The white solid was filtered, washed with acetone(2×15 mL) and dried in a vacuum oven at 50° C., affording the titlecompound, 0.637 g (51%). ¹H NMR (DMSO, 300 MHz) δ ppm 8.51 (s (broad),2H), 7.26 (t, 1H, J=8.1 Hz), 7.01 (m, 2H), 6.87 (dd, 1H, J=1.9 Hz, 7.3Hz), 6.29 (d, 1H, J=3.5 Hz), 4.84 (d, 1H, J=3.2 Hz), 3.74 (s, 3H), 3.16(m, 2H), 2.63 (t, 2H, J=6.8 Hz), 2.13 (m, 1H), 2.01 (m, 2H), 1.80 (m,2H), 1.51 (m, 3H), 0.92 (m, 1H), 0.71 (m, 1H). ¹³C (DMSO, 75 MHz) δ ppm159.37, 142.03, 129.62, 120.97, 114.51, 113.76, 72.89, 72.54, 55.96,50.10, 42.78, 31.96, 31.56, 25.35, 25.20, 23.29. ES-MS 342 (M−1).

Preparation of 3-{[(1S)-1-methylhexyl]amino}-1-propanesulfonic acid(Compound PS)

To a solution of (S)-(+)-2-aminoheptane (5.19 g, 45.0 mmol) in Acetone(25 mL) and Toluene (25 mL) was added 1,3-propane sultone (5.23 g, 42.9mmol). The solution was stirred at reflux for 4 hours. The reactionmixture was cooled to room temperature. The solid material was collectedby filtration and washed with acetone (2×20 mL). The solid was suspendedin EtOH (40 mL). The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature, the solid material was collectedby filtration, washed with acetone (2×20 mL) and dried in a vacuum ovenat 50° C., affording the title compound, 6.77 g (63%). ¹H NMR (D₂O, 300MHz) δ ppm 3.08 (m, 3H), 2.84 (t, 2H, J=7.3 Hz), 1.94 (m, 2H), 1.55 (m,1H), 1.38 (m, 1H), 1.18 (m, 9H), 0.70 (m, 3H). ¹³C (D₂O, 75 MHz) δ ppm55.00, 48.27, 43.59, 32.70, 31.06, 24.48, 22.17, 21.93, 15.79, 13.70.[α]_(D)=−6.4° (c=0.0020 in water), ES-MS 236 (M−1).

Preparation of 3-{[(1S)-1-methylheptyl]amino}-1-propanesulfonic acid(Compound PT)

To a solution of (S)-(+)-2-aminooctane (5.50 g, 42.5 mmol) in Acetone(25 mL) and Toluene (25 mL) was added 1,3-propane sultone (4.95 g, 40.5mmol). The solution was stirred at reflux for 4 hours. The reactionmixture was cooled to room temperature. The solid material was collectedby filtration and washed with acetone (2×20 mL). The solid was suspendedin EtOH (40 mL). The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature, the solid material was collectedby filtration, washed with acetone (2×20 mL) and dried in a vacuum ovenat 50° C., affording the title compound, 6.11 g (57%). ¹H NMR (D₂O, 300MHz) δ ppm 3.08 (m, 3H), 2.84 (t, 2H, J=7.3 Hz), 1.94 (m, 2H), 1.56 (m,1H), 1.39 (m, 1H), 1.16 (m, 11H), 0.70 (m, 3H). ¹³C (D₂O, 75 MHz) δ ppm55.00, 48.27, 43.59, 32.74, 31.19, 28.49, 24.75, 22.33, 21.93, 15.80,13.79. [α]_(D)=−6.5° (c=0.0031 in water), ES-MS 250 (M−1).

Preparation of 3-(1-propylamino)-1-propanesulfonic acid (Compound PU)

A mixture of propylamine (1.20 g, 20 mmo), 1,3-propane sultone (9.5 mLof 2.0 M in acetone) and toluene (7 mL) was heated to 50° C. for 3 h.The solid from the brownish suspension was collected by filtration,rinsed with acetone (2×5 mL) and dried 1 hour in vacuo (1.59 g). Thesolid was suspended in ethanol (10 mL) and the suspension was heated toreflux. Water (0.7 mL) was added and the mixture turned to a clearsolution. The mixture was then cooled with an ice/water bath. The solidwas collected by filtration, rinsed with acetone (2×5 mL) and dried 3days at 60° C. in the vacuum oven. The title compound was obtained as afine white powder 1.36 g, 7.56 mmol, 39%; ¹H NMR (500 MHz, D₂O) δ 0.97(t, J=7.6 Hz, 3H), 1.69 (hex, J=7.5 Hz, 2H), 2.12 (qt, J=7.6 Hz, 2H),3.01 (qt, J=7.3 Hz, 4H), 3.19 (t, J=7.8 Hz, 2H); ¹³C NMR (125 MHz, D₂O)δ 10.3, 19.3, 21.4, 46.2, 48.0, 49.4; ES-MS 180 (M−H)

Preparation of 3-{[(1R)-1-methylheptyl]amino}-1-propanesulfonic acid(Compound PV)

To a solution of (R)-(−)-2-aminooctane (5.00 g, 38.7 mmol) in acetone(25 mL) and toluene (25 mL) was added 1,3-propane sultone (4.50 g, 36.8mmol). The solution was stirred at reflux for 4 hours. The reactionmixture was cooled to room temperature. The solid material was collectedby filtration and washed with acetone (2×20 mL). The solid was suspendedin EtOH (40 mL). The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature, the solid material was collectedby filtration, washed with acetone (2×20 mL) and dried in a vacuum ovenat 50° C., affording the title compound, 5.72 g (56%). ¹H NMR (D₂O, 300MHz) δ ppm 3.09 (m, 3H), 2.84 (t, 2H, J=7.3 Hz), 1.95 (m, 2H), 1.56 (m,1H), 1.38 (m, 1H), 1.15 (m, 11H), 0.70 (m, 3H). ¹³C (D₂O, 75 MHz) δ ppm55.00, 48.26, 43.59, 32.74, 31.19, 28.48, 24.74, 22.31, 21.91, 15.79,13.78. [α]_(D)=+6.8° (c=0.0024 in water), ES-MS 250 (M−1).

Preparation of (R)-3-{[1-methylhexyl]amino}-1-propanesulfonic acid(Compound PW)

To a solution of (R)-(−)-2-aminoheptane (5.0 g, 43.4 mmol) in acetone(25 mL) and toluene (25 mL) was added 1,3-propane sultone (5.04 g, 41.3mmol). The solution was stirred at reflux for 4 hours. The reactionmixture was cooled to room temperature. The solid material was collectedby filtration and washed with acetone (2×20 mL). The solid was suspendedin EtOH (40 mL). The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature, the solid material was collectedby filtration, washed with acetone (2×20 mL) and dried in a vacuum ovenat 50° C., affording the title compound, 6.77 g (63%). ¹H NMR (D₂O, 300MHz) δ ppm 3.15 (m, 1H), 3.04 (m, 2H), 2.85 (t, 2H, J=7.3 Hz), 1.96 (m,2H), 1.57 (m, 1H), 1.39 (m, 1H), 1.19 (m, 9H), 0.72 (m, 3H). ¹³C (D₂O,75 MHz) δ ppm 54.84, 48.08, 43.38, 32.45, 30.80, 24.18, 21.88, 21.63,15.49, 13.35. [α]_(D)=+6.4° (c=0.0023 in water), ES-MS 236 (M−1).

Preparation of 3-[(3-oxocyclohex-1-en-1-yl)amino]-1-propanesulfonic acid(Compound PX)

A solution of 1,3-propane sultone (1.0 M in MeCN, 5.00 mL) was added toa solution of 3-amino-2-cyclohexenone (0.5558 g, 5.00 mmol) in a mixtureof MeCN (5 mL) and DMF (1.0 mL). The mixture was heated to 85° C. for 3hours on the Radley caroussel. The suspension was cooled to roomtemperature. The solid was collected by suction filtration, rinsed withacetone (2×5 mL). The solid was dried 18 hours at 60° C. in the vacuumoven. The title compound was obtained as a fine white solid (0.64 g,2.74 mmol, 55%). ¹H NMR (500 MHz, DMSO-d6) δ 2.01 (qt, J=6.5 Hz, 2H),2.22 (qt, J=6.8 Hz, 2H), 2.57 (t, J=6.3 Hz, 2H), 2.74 (t, J=6.6 Hz, 2H),3.05 (t, J=7.6 Hz, 2H), 4.28 (t, J=6.3 Hz, 2H), 5.85 (s, 1H); ¹³C NMR(125 MHz, DMSO-d6) δ 20.2, 23.7, 28.8, 28.9, 47.5, 68.7, 95.2, 183.4,186.2; ES-MS 340, 232 (M−H).

Preparation of 3-(1-butylamino)-1-propanesulfonic acid (Compound PY)

At reflux, a solution of 1,3-propane sultone (2.45 g, 20 mmol) intoluene (20 mL) was added dropwise over a 10 minutes period to asolution of to a solution of butylamine (1.46 g, 20 mmol) in acetone (20mL). The mixture was heated to reflux for 2 hours then was cooled toroom temperature. The solid was collected by suction filtration, rinsedwith acetone (2×5 mL). The solid was dried 1 hour at 60° C. in thevacuum oven (1.96 g). The solid was recrystallized in ethanol (20 mL).The mixture was left to cool to room temperature without stirring. Thesolid was collected by suction filtration, rinsed with acetone (2×5 mL).The solid was dried 18 hours at 60° C. in the vacuum oven. The titlecompound was obtained as a white solid (1.55 g, 7.94 mmol, 40%). ¹H NMR(500 MHz, D₂O) δ 0.92 (t, J=7.3 Hz, 3H), 1.38 (hex, J=7.5 Hz, 2H), 1.65(qt, J=7.7 Hz, 2H), 2.12 (qt, J=7.6 Hz, 4H), 3.00 (t, J=7.3 Hz, 2H),3.05 (t, J=7.6 Hz, 2H), 3.18 (t, J=7.8 Hz, 2H); ¹³C NMR (125 MHz, D₂O) δ12.9, 19.3, 21.5, 27.7, 46.3, 47.6, 48.1; ES-MS 194 (M−H).

Preparation of 3-[benzyl(tert-butyl)amino]-1-propanesulfonic acid(Compound PZ)

A mixture of 1,3-propane sultone (12.30 g, 100 mmol), toluene (20 mL)N-benzyl-N-tert-butylamine (16.33 g, 100 mmol) in cyclohexanenone (100mL) was heated to reflux for 2 hours, diluted with toluene then cooledto room temperature. The mixture was stirred overnight at roomtemperature then diluted with acetone (100 mL). The solid was collectedby suction filtration, rinsed with acetone (2×50 mL). The solid wasdried 6 hours at 60° C. in the vacuum oven (23 g). The solid wasrecrystallized in methanol (150 mL) and water (38 mL). The mixture wasleft to cool to room temperature. The solid was collected by suctionfiltration, rinsed with methanol (2×5 mL). The solid was dried 18 hoursat 60° C. in the vacuum oven. The title compound was obtained as a whitesolid 12.75 g in the first crop. The mother liquor was concentrated to athick paste and refluxed with ethanol (80 mL) for 30 minutes. The solidwas collected by filtration, rinsed with ethanol (2×20 mL) and was dried18 hours at 60° C. in the vacuum oven to afford a second crop of 6.42 gfor a total yield: 19.17 g, 67.17 mmol, 67%. ¹H NMR (500 MHz, D₂O) δ0.99 (m, 1H), 1.54 (s, 9H), 1.80 (m, 1H), 2.53 (m, 1H), 2.64 (m, 1H),3.21 (m, 1H), 3.59 (m, 1H), 4.08 (br d, J=12.7 Hz, 3H), 4.71 (br d,J=12.7 Hz, 3H), 7.50-7.58 (m, 5H); ¹³C NMR (125 MHz, D₂O) δ22.8, 24.3,47.8, 49.3, 66.3, 129.7, 130.1, 130.4, 131.3; ES-MS 284 (M−H).

Preparation of (R)-3-{[1-(3-methoxyphenyl)ethyl]amino}-1-propanesulfonicacid (Compound QA)

To a solution of (1R)-(−)-1-(3-methoxyphenyl)ethylamine (5.0 g, 33.1mmol) in acetonitrile (30 mL) and toluene (10 mL) was added 1,3-propanesultone (3.85 g, 31.5 mmol). The solution was stirred at reflux for 4hours. The reaction mixture was cooled to room temperature. The solidmaterial was collected by filtration and washed with acetone (2×20 mL).The solid was suspended in EtOH (40 mL). The suspension was stirred atreflux for 1 hour. The mixture was cooled to room temperature, the solidmaterial was collected by filtration, washed with acetone (2×20 mL) anddried in a vacuum oven at 50° C., affording the title compound, 7.98 g(91%). ¹H NMR (D₂O, 500 MHz) δ ppm 7.298 (td, 1H, J=1.0 Hz, 7.8 Hz),6.92 (m, 3H), 4.23 (q, 1H, J=6.8 Hz), 3.69 (s, 3H), 2.95 (m, 1H), 2.78(m, 3H), 1.92 (m, 2H), 1.50 (d, 3H, J=6.8 Hz). ¹³C (D₂O, 125 MHz) δ ppm159.69, 137.59, 130.99, 120.23, 115.36, 113.38, 58.53, 55.64, 48.08,44.50, 21.52, 18.42. [α]_(D)=+23.7° (c=0.0036 in water), ES-MS 272(M−1).

Preparation of 3-[(1,1-dimethylbut-3-enyl)amino]-1-propanesulfonic acid(Compound QB)

A 100-mL., three-necked, round-bottomed flask equipped with stir-bar,dropping funnel, and low-temperature thermometer is charged with 2.03 g.(15.8 mmole) of 2,2-dimethylpentenoic acid and 15 mL of acetone. Themixture is stirred, and 2.45 mL. (17.4 mmole) of triethylamine is addedover 5 minutes. The solution is chilled to −5 to 0° in an ice-salt bath,and 1.67 mL. (17.4 mmole) of ethyl chlorocarbonate in 5 mL of acetone isadded slowly (25 minutes), maintaining the temperature between −5 to 0°.After the addition is complete, the cold mixture is stirred for anadditional 15 minutes. A solution of 2.05 g. (31.6 mmole) of sodiumazide in 8 mL of water is added over a 25-minute period while thetemperature is kept at −5 to 0°. The mixture is stirred for 30 minuteslonger at this temperature, poured into 75 ml. of ice water, and shakenwith four 25-ml. portions of toluene. The combined toluene extracts aredried over anhydrous magnesium sulfate and transferred to a 250 mL,three-necked, round-bottomed flask equipped with a two-necked,Claisen-type adapter, stirrer, and reflux condenser. The stirredsolution is heated cautiously under reflux for 1 hour (nitrogenevolution is observed initially). The amine hydrochloride solution wasconcentrated to dryness. The amine hydrochloride was dissolved in aminimum of hot methanol (4 mL) and was poured into ether (20 mL). Theamine was collected by filtration and dried in vacuo. Proton NMRindicated a purity of about 97% (trace of triethylamine hydrochloride)(1.27 g, 9. mmol, 59%). The amine was dissolved in water and thesolution was made to pH 12 with a saturated potassium carbonatesolution. The amine was extracted with MTBK (4×5 mL), dried over sodiumsulfate, rinsed with toluene (5 mL). The solution obtained was used assuch in the next step. ¹H NMR (500 MHz, D₂O) δ 1.34 (s, 6H), 1.38 (hex,J=7.5 Hz, 2H), 2.39 (d, J=7.3 Hz, 2H), 5.23-5.29 (m, 2H), 5.84-5.92 (m,1H); ¹³C NMR (125 MHz, D₂O) δ 24.7, 44.0, 54.1, 121.2, 131.3

To the solution of 1,1-dimethylbut-3-enylamine (9 mmol) in MTBK (20mL)/toluene (5 mL) was added 1,3-propane sultone (0.8 mL, 9 mmol). Themixture was heated to gentle reflux for 5 hours then was cooled to roomtemperature. The solid was collected by suction filtration, rinsed withacetone (2×5 mL). The solid was dried 60 hours at 60° C. in the vacuumoven. The title compound was obtained as a beige solid (0.83 g, 3.75mmol, 42% from the amine hydrochloride, 24% overall). ¹H NMR (500 MHz,D₂O) δ 1.34 (s 6H), 2.08 (qt, J=7.6 Hz, 2H), 2.44 (d, J=7.3 Hz, 4H),3.00 (t, J=7.3 Hz, 2H), 3.19 (t, J=7.8 Hz, 2H), 5.28-5.31 (m, 2H),5.82-5.90 (m. 2H); ¹³C NMR (125 MHz, D₂O) δ 12.9, 19.3, 21.5, 27.7,46.3, 47.6, 48.1; ES-MS 194 (M−H)

Preparation of 3-[(4-methylbenzyl)atnino]-1-propanesulfonic acid(Compound QC)

To a solution of 4-methylbenzylamine (5.0 g, 41.3 mmol) in acetonitrile(35 mL) and toluene (15 mL) was added 1,3-propane sultone (4.80 g, 39.3mmol). The solution was stirred at reflux for 4 hours. The reactionmixture was cooled to room temperature. The solid material was collectedby filtration and washed with acetone (2×20 mL). The solid was suspendedin EtOH (40 mL). The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature, the solid material was collectedby filtration, washed with acetone (2×20 mL) and dried in a vacuum ovenat 50° C., affording the title compound, 8.62 g (90%). ¹H NMR (D₂O, 500MHz) δ ppm 7.20 (d, 2H, J=7.8 Hz), 7.17 (d, 2H, J=7.8 Hz), 4.04 (s, 2H),3.05 (t, 2H, J=7.8 Hz), 2.82 (t, 2H, J=7.3 Hz), 2.20 (s, 3H), 1.97 (m,2H). ¹³C (D₂O, 125 MHz) δ ppm 140.37, 129.99, 127.77, 51.00, 48.09,45.78, 21.43, 20.48. ES-MS 242 (M−1).

Preparation of3-{[2-(4-methoxyphenyl)-2-oxoethyl]amino}-1-propanesulfonic acid(Compound QD)

2-Amino-4-methoxyacetophenone hydrochloride (2.5 g, 12.4 mmol) wastreated with a saturated solution of K₂CO₃ (65 mL) and EtOAc (3×65 mL)was added. The organic extracts were combined, dried with Na₂SO₄,filtered, evaporated under reduced pressure and dried in vacuo.

To a solution of 2-amino-4-methoxyacetophenone (12.4 mmol) in 25%Toluene/Acetonitrile (15 mL) was added 1,3-propane sultone solution(1.34 g, 11.0 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×20 mL). The beigesolid was dissolved with heating in 50% MeOH/Water (200 mL) before DowexMarathon C ion exchange resin (strongly acidic) was added. Thesuspension was stirred at room temperature for 15 minutes. The resin wasfiltered and washed with 50% MeOH/Water (2×15 mL). The filtrate wasevaporated. The solid was suspended in acetone before it was collectedby filtration, washed with acetone (2×20 mL) and dried in a vacuum oven(50° C.), affording the title compound, 1.84 g (58%). ¹H NMR (DMSO, 500MHz) ppm 9.06 (s (broad), 1H)), 7.98 (d, 2H, J=8.8 Hz), 7.12 (d, 2H,J=8.8 Hz), 4.72 (m, 2H), 3.87 (s, 3H), 3.13 (m, 2H), 2.62 (t, 2H, J=6.6Hz), 2.02 (m, 2H). ¹³C (D₂O, 75 MHz) δ ppm 191.04, 164.69, 131.24,127.09, 114.97, 56.64, 52.82, 49.95, 47.80, 22.63. ES-MS 286 (M−1).

Preparation of 3-[(1,1-dimethylprop-2-enyl)amino]-1-propanesulfonic acid(Compound QE)

A dry, 250-ml., three-necked flask is equipped with a magnetic stirringbar, a pressure-equalizing dropping funnel, a thermometer, and anitrogen inlet tube. The apparatus was flushed with nitrogen and chargedwith 410 mg. (0.0103 mole) of sodium hydride dispersed in mineral oiland 15 mL of hexane. The suspension was stirred, and the hydride wasallowed to settle. The hexane was removed with a long dropping pipette,60 mL of anhydrous diethyl ether was added, and a solution of 8.55 g.(0.0993 mole) of 3-methyl-2-butenol in 15 mL of anhydrous ether wasadded over 5 minutes. After the evolution of hydrogen ceases, thereaction mixture was stirred for an additional 15 minutes. The clearsolution was then cooled to between −10 and 0° in an ice-salt bath.Trichloroacetonitrile (10.0 ml., 14.4 g., 0.0996 mole) was addeddropwise to the stirred solution, while the reaction temperature wasmaintained below 0°. The addition was completed within 15 minutes, andthe reaction mixture was allowed to warm to room temperature. The lightamber mixture was poured into a 250-ml., round-bottomed flask, and theether was removed with a rotary evaporator. Hexane [150 ml., containing0.4 ml. (0.01 mole) of methanol] was added, the mixture was shakenvigorously for 1 minute, and a small amount of dark, insoluble materialis removed by gravity filtration. The residue was washed two times withhexane (50 ml. total), and the combined filtrate was concentrated with arotary evaporator

A 500-ml., round-bottomed flask was charged with imidate and 300 mL ofxylene. The solution was refluxed for 8 hours. After cooling to roomtemperature the dark xylene solution was filtered through a short columnpacked with silica gel and toluene. The column was eluted with anadditional 250 mL of toluene, and the combined light yellow eluant isconcentrated with a rotary evaporator.

A 500-mL round-bottomed was charged with 9.0 g. (0.030 mole) of thecrude amide, 160 mL of ethanol, and 150 mL of aqueous 6 N sodiumhydroxide. The air was replaced with nitrogen, and the solution wasstirred at room temperature for 40 hours. Ether (300 mL) was added, theorganic layer was separated, and the aqueous layer was washed twice with50 mL of ether.

After extraction in ether (5×100 mL), the crude amine was back extractedin 6N HCl (2×20 mL). The combined organic extract were washed with ether(1×100 mL). The acid solution was adjusted to pH 12 with 50% NaOH. Thebasic aqueous layer was extracted with ether (2×50 mL). A solution of 2MHCl in ether (100 mL) was added and the amine hydrochloride solution wasconcentrated to dryness. The amine hydrochloride was dissolved in aminimum of hot methanol (10 mL) and was poured into ether (100 mL). Theamine was collected by filtration and dried in vacuo. The pure amindehydrochloride was obtained in two crops (6.92 g, 56.9. mmol, 57%). Theamine was dissolved in water and the solution was made to pH 12 with 50%NaOH. The amine was extracted with toluene (3×13 mL0 then MTBK (1×10mL), dried over potassium carbonate (20 minutes), rinsed with acetone (5mL). ¹H NMR (500 MHz, D₂O) δ 1.34 (s 6H), 5.30 (d, J=10.7 Hz, 1H), 5.31(d, J=17.6 Hz, 1H), 5.98 (dd, J=17.6 and 11.2 Hz, 1H); ¹³C NMR (125 MHz,D₂O) 25.1, 54.9, 115.5, 139.3

Preparation of 3-[(1-Carbamoyl-1-ethyl)propylamino]-1-propanesulfonicacid (Compound QF)

To a 250 mL 1 neck flask containing 30% NH₄OH (120 mL) was added NaCN(15.34 g, 0.31 mol) and NH₄Cl (19.75 g, 0.37 mol) with vigorousstirring. The corresponding ketone was added dropwise within 20 minutesat room temperature. The mixture was stirred for 3 days at roomtemperature followed by extraction with dichloromethane (50 mL). Theorganic layer was separated and dried over anhydrous sodium sulfate for2 hours. The sodium sulfate was removed by filtration, the solvent wasremoved under reduced pressure to yield the crude aminonitrile. Thedesired material was obtained as an light brown oil (colorless oil, 89%crude yield). ¹H NMR (500 MHz, DMSO-d6) δ 0.95 (t, J=7.6 Hz, 6H), 1.52(hex, J=7.2 Hz, 2H), 1.61 (hex, J=7.3 Hz, 2H), 2.39 (br s, 2H); ¹³C NMR(125 MHz, DMSO-d6) δ 8.3, 31.9, 54.6, 124.2

To 10 g of concentrated sulfuric acid stirred in an ice cooled waterbath was added dropwise a solution of the aminonitrille (41 mmol) in 30mL CH₂Cl₂, maintaining the internal temperature at 15° C. Then the bathwas removed and the mixture heated to 40° C. for 1 hour. The mixture wascooled in ac ice bath and poured onto 200 g of crushed ice. The mixturewas made pH 7-8 with 28% aqueous NH₃ and extracted with EtOAc (3×100mL). The extracts were collected, dried (MgSO₄), and evaporated todryness. The crude solid was recrystallized in EtOAc/Hex. The desiredmaterial was obtained as a white foamy solid 0.941 g, 7.23 mmol, 6%. ¹HNMR (500 MHz, DMSO-d6) δ 0.77 (t, J=7.6 Hz, 6H), 1.29-1.36 (m, 2H),1.57-1.65 (m, 2H), 6.95 (br s, 1H), 7.23 (br s, 1H); ¹³C NMR (125 MHz,DMSO-d6) δ 8.2, 32.5, 60.6, 178.2

One equivalent of 1,3-propane was added to a solution of2-amino-2-ethylpropaneamide (0.941 g, 7.23 mmol). A paste was obtainedafter the reaction. The paste was dissolved in water and washed withethyl acetate. The sodium salt was prepared with 1N NaOH and thesolution was concentrated to dryness. The crude product was purified bypreparative RP-HPLC (Delta Prep pack cartridge C18, 215 nm, 50 mL/min,0% to 30% McCN in water containing 0.01% TFA). After freeze-drying, thetitle compound was obtained as a fine white solid (0.3700 g, 1.47 mmol,20%). ¹H NMR (300 MHz, D₂O) δ 0.89 (t, J=7.5 Hz, 6H), 1.83 (q, J=7.3 Hz,4H), 2.05 (qt, J=7.4 Hz, 4H), 2.84-2.86 (m, 2H), 2.99 (t, J=7.3 Hz, 2H);¹³C NMR (75 MHz, D₂O) δ 7.4, 23.5, 26.0, 41.5, 48.9, 68.3, 176.9; ES-MS251 (M−H).

Preparation of 3-[(4-tert-butylbenzyl)amino]-1-propanesulfonic acid(Compound QG)

To a solution of 4-tert-butylbenzylamine (5.0 g, 30.6 mmol) in 25%Toluene/Acetonitrile (30 mL) was added 1,3-propane sultone solution(3.56 g, 29.1 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×20 mL). The solid wassuspended in EtOH (50 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×20 mL) and dried in avacuum oven (50° C.), affording the title compound, 7.58 g (91%). ¹H NMR(DMSO, 500 MHz) δ ppm 8.95 (s (broad), 1H), 7.44 (d, 2H, J=8.3 Hz), 7.38(d, 2H, J=8.3 Hz), 4.07 (s, 2H), 3.08 (t, 2H, J=6.3 Hz), 2.63 (t, 2H,J=6.5 Hz), 1.96 (m, 2H), 1.27 (s, 9H). ¹³C (DMSO, 75 MHz) δ ppm 154.21,132.32, 132.05, 128.31, 52.77, 52.32, 49.89, 34.17, 24.81. ES-MS 284(M−1).

Preparation of 3-{[1-(3-methoxyphenyl)propyl]amino}-1-propanesulfonicacid (Compound QH)

To a 0° C. solution of m-Anisaldehyde (27 mL, 22 mmol) in anhydroustetrahydrofuran (THF, 5 mL) was added dropwise lithiumbis(trimethylsilyl)amide (1M solution in THF, 26 mL, 26 mmol). Thesolution was stirred at 0° C. for 20 minutes before ethylmagnesiumbromide (1M solution in THF, 28 mL, 28 mmol) via syringe. The reactionmixture was stirred at reflux for 24 hours. After cooling to roomtemperature, the reaction mixture was poured into a saturated solutionof NH₄Cl (50 mL). The mixture was extracted with EtOAc (3×75 mL). Theorganic extracts were combined and concentrated under reduced pressure.The residue was stirred with 3M HCl (40 mL) for 30 minutes. Theresulting mixture was extracted with EtOAc (3×40 mL). The combinedorganic extracts were dried with Na₂SO₄, filtered, evaporated and driedin vacuo, affording 1-(3-methoxyphenyl)-1-propanamine (2.47 g, 67%).

To a solution of 1-(3-methoxyphenyl)-1-propanamine (2.45 g, 14.8 mmol)in 25% toluene/acetonitrile (15 mL) was added 1,3-propane sultonesolution (1.72 g, 14.1 mmol). The solution was stirred at reflux for 4hours. The reaction mixture was cooled to room temperature. The solidmaterial was collected by filtration and washed with acetone (2×20 mL).The solid was suspended in EtOH (20 mL). The suspension was stirred atreflux for 1 hour. The mixture was cooled to room temperature, the solidmaterial was collected by filtration, washed with acetone (2×20 mL) anddried in a vacuum oven (50° C.), affording the title compound, 3.16 g(78%). ¹H NMR (D₂O, 500 MHz) δ ppm, 7.28 (t, 1H, J=7.9 Hz), 6.90 (m,3H), 3.98 (m, 1H), 3.69 (m, 3H), 2.89 (m, 1H), 2.74 (m, 3H), 1.89 (m,4H), 0.63 (t, 2H, J=7.3 Hz). ¹³C (D₂O, 75 MHz) δ ppm 159.47, 135.45,130.81, 120.84, 115.32, 113.87, 64.50, 55.72, 48.23, 44.73, 26.14,21.72, 9.80. ES-MS 286 (M−1).

Preparation of3-{[2-(2-hydroxyphenyl)-1,1-dimethylethyl]amino}propane-1-sulfonic acid(Compound QI)

The benzyl alcohol (1.2 g, 10 mmol), the tetrabutylammonium fluoride (5mL, 5 mmol) and the nitro compound (1.78 g, 20 mmol) were placed in asealed tube and heated at 130° C. for 15 hours. The reaction was cooledand diluted with EtOAc. The resulting solution was washed with water,dried and concentrated to yield a dark oil. Chromatography over silicaeluting with Hex:EA 80:20 gave a yellowish solid 0.48 g, 25% yield.

To a stirred solution of the nitro (800 mg, 4.12 mmol) in methanol (20mL) was added a small spatula of Raney-Ni in water. The suspension washydrogenated under atmospheric pressure of hydrogen for 15 hours (TLCindicates complete consumption of the starting material) then filteredon celite and concentrated under reduced pressure. The correspondingamine was used as such in the next step.

To a stirred solution of the amine (420 mg, 2.58 mmol) in THF (5 mL) wasadded 1,3-propane sultone (614 mg, 5.02 mmol). The reaction mixture wasstirred at reflux for 15 hours then cooled to room temperature. Thesolid was collected by filtration and was washed with THF. The solid wassuspended in EtOH (10 mL) and stirred at reflux for 1 hour. Thesuspension was then cooled to room temperature. The solid was collectedby filtration, washed with ethanol and dried under high vacuum to affordthe title compound, 75 mg (10% yield). ¹H NMR (500 MHz, DMSO-d₆) δ 1.17(s, 6H), 2.00 (m, 2H), 2.64 (m, 2H), 2.85 (s, 2H), 3.12 (m, 2H),6.70-6.82 (m, 2H), 7.10 (m, 2H), 8.49 (bs, 2H), 9.64 (s, 1H, OH). ¹³CNMR (125 MHz, DMSO-d₆) δ 23.42, 23.73, 37.98, 41.51, 49.89, 60.54,115.99, 119.44, 122.09, 128.87, 133.14, 156.30. ES-MS 286 (M−1).

Preparation of 3-[(1-methyl-1-thien-2-ylethyl)amino]-1-propanesulfonicacid (Compound QJ)

CeCl₃-7H₂O was dried at 140° C.-150° C. for 15 hours. To this solid wasadded THF (80 mL), and after stirring for 30 minutes the suspension wascooled to −78° C. and to it was added MeLi. After stirring for 30minutes, 2-thiophencarbonotrile was added dropwise, and the reaction wasstirred at −78° C. to −35° C. for 3 hours. Concentrated aqueous NH₃ wasadded (25 mL) and the mixture was warmed to room temperature andfiltered through celite. The filtrate was extracted with EtOAc dried(Na2SO4) and concentrated. Column using CH₂Cl₂/MeOH (95/05) allowed theisolation of the desired product, 600 mg (23% yield).

To a stirred solution of the amine (500 mg, 3.5 mmol) in pinacolone (7mL) was added 1,3-propane sultone (427 mg, 3.5 mmol). The reactionmixture was stirred at reflux for 4 hours then cooled to roomtemperature. The solid was collected by filtration and was washed withTHF. The solid was suspended in EtOH (10 mL) and stirred at reflux for 1hour. The suspension was then cooled to room temperature. The solid wascollected by filtration, washed with ethanol and dried under high vacuumto afford the title compound, 800 mg (87%). ¹H NMR (500 MHz, D₂O) δ 1.70(s, 6H), 1.85 (m, 2H), 2.75 (t, J=7.0 Hz, 2H), 2.84 (t, J=7.0 Hz, 2H),7.00 (m, 1H), 7.16 (m, 1H), 7.40 (m, 1H) ¹³C NMR (125 MHz, DMSO-d₆) δ21.75, 25.82, 41.07, 48.07, 59.52, 127.67, 127.90, 127.95. ES-MS 262(M−1).

Preparation of 3-{[4-(methylsulfonyl)benzyl]amino}-1-propanesulfonicacid (Compound QK)

4-methylsulfonylbenzylamine hydrochloride (2.5 g, 11.8 mmol) was treatedwith a saturated solution of K₂CO₃ (40 mL) and EtOAc (3×40 mL) wasadded. The organic extracts were combined, dried with Na₂SO₄, filtered,evaporated under reduced pressure and dried in vacuo.

To a solution of 4-methylsulfonylbenzylamine (2.11 g, 11.4 mmol) in 25%Toluene/Acetonitrile (15 mL) was added 1,3-propane sultone solution(1.35 g, 10.8 mmol). The solution was stirred at reflux for 4 hours. Thereaction mixture was cooled to room temperature. The solid material wascollected by filtration and washed with acetone (2×20 mL). The solid wassuspended in EtOH (20 mL). The suspension was stirred at reflux for 1hour. The mixture was cooled to room temperature, the solid material wascollected by filtration, washed with acetone (2×20 mL) and dried in avacuum oven (50° C.), affording the title compound, 3.10 g (91%). ¹H NMR(D₂O, 300 MHz) δ ppm 7.87 (dd, 2H, J=1.9 Hz, 6.6 Hz), 7.57 (dd, 2H,J=1.9 Hz, 6.6 Hz), 4.22 (s, 2H), 3.12 (m, 3H), 2.84 (t, 2H, J=7.3 Hz),2.00 (m, 2H). ¹³C (D₂O, 75 MHz) δ ppm 139.83, 137.22, 130.92, 128.02,50.59, 48.17, 46.50, 43.48, 21.69. ES-MS 306 (M−1).

Preparation of (R)-(+)-3-[1-(4-nitrophenyl)ethylamino]-1-propanesulfonicacid (Compound QL)

A solution of 1,3-propane sultone (1M, 5.0 mL) in acetonitrille wasadded to a solution of (R)-(+)-1-(4-nitrophenyl)ethylamine (0.8185 g,4.93 mmol) in toluene (10 mL). The mixture was heated to reflux for 24hours then cooled to 0° C. The solid was collected by suctionfiltration, rinsed with acetone (2×5 mL). The solid was dried 4 hours at60° C. in the vacuum oven. The title compound was obtained as a finewhite solid (1.14 g, 3.95 mmol, 80%). ¹H NMR (300 MHz, DMSO-d6) δ 1.55(d, J=6.7 Hz, 3H), 1.97 (qt, J=6.5 Hz, 2H), 2.62 (t, J=6.4 Hz, 2H), 2.84(br s, 1H), 3.05 (br s, 2H), 4.58 (br s, 1H), 7.77 (d, J=8.8 Hz, 1H),8.31 (d, J=8.8 Hz, 2H), 9.21 (br s, 1H), 9.37 (br s, 1H); ¹³C NMR (75MHz, DMSO-d6) 19.0, 22.0, 45.2, 49.0, 56.0, 123.8, 128.8, 147.4, 147.4;ES-MS 287 (M−H); [α]_(D)=+37.1°±0.1, c=0.005512, 0.1 N NaOH

Preparation of 3-[(1,1-diethylprop-2-enyl)amino]-1-propanesulfonic acid(Compound QM)

A solution of triethyl phosphonoacetate (11.4 g, 50.8 mmol) in ether (50mL) was added dropwise to a suspension of sodium hydride (60% in oil,2.20 g, 55 mmol) in ether (100 mL) at 0° C. The mixture was stirred 1hour at room temperature. The clear solution was cooled again to 0° C.and a solution of 3-pentanone (5.85 mL, 55 mmol) in ether (20 mL) wasadded dropwise over 20 minutes. The mixture was then heated overnight atreflux. The liquid from the flask was decanted to leave the phosphatesodium salt behind. The solid was rinsed with ether (3×20 mL). Thecombined organic phase was rinsed with water (1×20 m), 1N NaOH (1×20 mL)and brine (1×20 mL). The organic layers was dried over sodium sulfateand the solution was concentrated to an oil under reduced pressure. Thecrude residue was flash chromatographied on a 70 g silica gel cartridgeon a Biotage system to afford the desire material as a clear oil (5.37g, 34.4 mmol, 68% yield, about 90% pure: some phosphonate). ¹H NMR (500MHz, CDCl₃) δ 1.069 (t, J=7.3 Hz, 3H), 1.073 (t, J=7.6 Hz, 3H), 2.190(q, J=7.5 Hz, 1H), 2.192 (q, J=7.5 Hz, 1H), 2.62 (q, J=7.6 Hz, 2H), 4.15(qt, J=7.2 Hz, 2H), 5.60 (s, 1H); ¹³C NMR (75 MHz, CDCl₃) δ 12.2, 13.2,14.5, 25.6, 30.9, 59.5, 113.5, 166.3, 167.0.

A solution of 3-Ethyl-2-pentenoic acid ethyl ester (5.30 g, 34 mmol) inether (40 mL) was added dropwise over 20 minutes to an ico-coldsuspension of LAH (1.5 g) in ether (150 g). The mixture was stirred for5 minutes at 0° C. The mixture was quenched with methanol then asaturated sodium tartrate solution was added. The layers were separatedand the organic layer was dried over sodium sulfate, concentrated todryness. The crude alcohol obtained was flashed on a 70 g silica gelcartridge (Bioatge) using 20 to 30% ether in hexane. The desired productwas obtained as a clear oil, 1.55 g, 14.5 g, 43%. ¹H NMR (500 MHz,CDCl₃) δ 0.96-1.03 (m, 6H), 1.36 (m, 1H), 2.07-2.12 (m, 4H), 4.17 (d,J=6.8 Hz, 2H), 5.36 (t, J=7.1 Hz, 1H); ¹³C NMR (75 MHz, CDCl₃) δ 12.6,13.9, 23.7, 29.2, 59.1, 121.3, 147.1.

A dry, 50-mL, three-necked flask was equipped with a magnetic stirringbar, a pressure-equalizing dropping funnel, a thermometer, and anitrogen inlet tube. The apparatus was flushed with nitrogen and chargedwith 55 mg of sodium hydride dispersed in mineral oil and 3 mL ofhexane. The suspension was stirred, and the hydride was allowed tosettle. The hexane was removed with a long dropping pipette, 8 mL ofanhydrous diethyl ether was added, and a solution of 8.55 g. (0.0993mole) of 3-ethyl-2-pentenol in 3 mL of anhydrous ether was added over 5minutes. After the evolution of hydrogen ceased, the reaction mixturewas stirred for an additional 15 minutes. The clear solution was cooledto −10° C. in an ice-salt bath. Trichloroacetonitrile (1.35 ml, 13.4mmole) was added dropwise to the stirred solution, while the reactiontemperature was maintained below 0°. Addition was completed within 15minutes, and the reaction mixture was allowed to warm to roomtemperature. The light amber mixture was poured into a 100 mL,round-bottomed flask, and the ether was removed with a rotaryevaporator. Hexane [20 mL, containing a drop of methanol] was added, themixture was shaken vigorously for 1 minute, and a small amount of dark,insoluble material was removed by gravity filtration. The residue iswashed two times with hexane (10 mL total), and the combined filtrate isconcentrated with a rotary evaporator.

A 100-mL, round-bottomed flask was charged with the imidate and 40 mL ofxylene. The solution is refluxed for 8 hours. After cooling to roomtemperature the dark xylene solution was filtered through a short columnpacked with silica gel (20 g) and toluene. The column was eluted with anadditional 40 mL of toluene, and the combined light yellow eluant wasconcentrated with a rotary evaporator.

To the crude product was added 60 ml. of ethanol and 60 ml. of aqueous 6N sodium hydroxide. The air was replaced with nitrogen, and the solutionwas stirred at room temperature for 40 hours. Ether (300 mL) is added,the organic layer is separated, and the aqueous layer is washed twicewith 50 mL of ether.

After extraction in ether (5×20 mL), the crude amine was back extractedin 6N HCl (2×20 mL). The combined organic extract were washed with ether(1×100 mL). The acid solution was adjusted to pH 12 with 50% NaOH. Thebasic aqueous layer was extracted with ether (2×50 mL). A solution of 2MHCl in ether (100 mL) was added and the amine hydrochloride solution wasconcentrated to dryness. The amine hydrochloride was dissolved in aminimum of hot methanol (4 mL) and was poured into ether (50 mL). Theamine was collected by filtration and dried in vacuo. The pure amindehydrochloride was obtained in two crops (0.67 g, 4.48 mmol, 34%). Theamine was dissolved in water and the solution was made to pH 12 with 50%NaOH. The amine was extracted with toluene (1×5 mL) then MTBK (2×5 mL),dried over potassium carbonate (20 minutes). NMR data on the aminehydrochloride: ¹H NMR (500 MHz, D₂O) δ 0.91 (t, J=7.6 Hz, 6H), 1.74-1.79(m, 4H), 5.22 (d, J=17.6 Hz, 1H), 5.40 (d, J=11.2 Hz, 1H), 5.85 (dd,J=18.1 and 11.2 Hz, 1H); ¹³C NMR (75 MHz, D₂O) δ 12.6, 13.9, 23.7, 29.2,59.1, 121.3, 147.1

The 1,3-propane sultone (0.40 mL, 4.5 mmol) was added to a solution ofto a solution of 1,1-diethylprop-2-enylamine (4.48 mmol) in 20%MTBK/toluene (15 mL). The mixture was heated to reflux for 20 hours thenwas cooled to 0° C. The solid was collected by suction filtration,rinsed with acetone (2×5 mL). The solid was dried 6 hours at 60° C. inthe vacuum oven. The title compound was obtained as a white solid (0.58g, 2.46 mmol, 55%, 5% overall). ¹H NMR (300 MHz, DMSO-d6) δ 0.82 (t,J=7.5 Hz, 6H), 1.69 (non, J=7.3 Hz, 4H), 1.99 (qt, J=6.4 Hz, 2H), 2.65(t, J=6.4 Hz, 2H), 2.87-2.90 (m, 2H), 5.35 (d, J=17.6 Hz, 1H), 5.48 (d,J=11.1 Hz, 1H), 5.75 (dd, J=17.6 and 11.1 Hz, 1H), 8.71 (br s, 2H); ¹³CNMR (75 MHz, DMSO-d6) δ 7.1, 22.1, 24.2, 41.2, 64.5, 119.7, 136.0; ES-MS234 (M−H).

Preparation of 4-(tert-butylamino)-1-phenyl-2-butanesulfonic acid(Compound QN)

To a −78° C. solution of 1,3-propane sultone (1 eq) in anhydroustetrahydrofuran (THF, 1.8 M) was added butyl lithium (2.5 M in hexanes,1.5 eq). The solution was stirred at −78° C. for 0.5 hours before benzylbromide (1 eq diluted with THF) was added via syringe pump over 0.5hours period. The reaction mixture was stirred at −78° C. for 2 hours.The reaction mixture was warmed up to 0° C. before water (100 mL) wasslowly added. The organic layer was recovered and diluted with EtOAc (20mL). The solution was dried with Na₂SO₄, filtered and evaporated underreduced pressure. The product was purified on a silica gel pad (90%Hex/EtOAc to 60% Hex/EtOAc) affording the resulting 1-Benzyl-1,3-propanesultone.

To a solution of 1-Benzyl-1,3-propane sultone (1 eq) in 25%Toluene/Acetonitrile (0.8 M) was added Tert-Butylamine (1.05 eq). Thesolution was stirred at reflux for 4 hours. The reaction mixture wascooled to room temperature. The solid material was collected byfiltration and washed with acetone (2×20 mL). The solid material wassuspended in EtOH. The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature, the solid material was collectedby filtration, washed with acetone (2×20 mL) and dried in a vacuum oven(50° C.), affording the title compound. ¹H NMR (D₂O, 500 MHz) δ ppm 7.23(m, 5H), 3.28 (dd, 1H, J=3.7 Hz, 13.9 Hz), 3.06 (m, 1H), 2.87 (m, 1H),2.58 (m, 2H), 1.90 (m, 1H), 1.76 (m, 1H), 1.06 (s. 9H). ¹³C (D₂O, 125MHz) δ ppm 138.09, 129.45, 129.13, 127.25, 59.41, 57.09, 39.49, 36.11,26.40, 24.90. ES-MS 284 (M−1).

Preparation of 1-(tert-butylamino)hex-5-ene-3-sulfonic acid (CompoundQO)

To a −48° C. solution of 1,3-propane sultone (5.0 g, 41 mmol) inanhydrous tetrahydrofuran (THF, 150 mL) was added butyl lithium (2.5 Min hexanes, 18 mL, 45 mmol). The solution was stirred at −78° C. for 30minutes before benzyl bromide (3.5 mL, 41 mmol) was added via syringepump over a 30 minute period. The reaction mixture was stirred at −48°C. for 2 hours. The reaction mixture was warmed up to 0° C. before water(100 mL) was slowly added. The organic layer was recovered and dilutedwith EtOAc (20 mL). The solution was dried with Na₂SO₄, filtered andevaporated under reduced pressure. The product was purified on a silicagel column (90% Hex/EtOAc to 70% Hex/EtOAc) affording the corresponding1-allyl-1,3-propane sultone (3.03 g, 46%).

To a solution of Tert-Butylamine (1.43 g, 19.6 mmol) in 25%Acetonitrile/Toluene (20 mL) was added 1-allyl-1,3-propane sultone (3.03g, 18.6 mmol) in 25% Acetonitrile/Toluene (5 mL). The solution wasstirred at reflux for 4 hours. The solution was stirred at reflux for 4hours. The reaction mixture was cooled to room temperature. The solidmaterial was collected by filtration and washed with acetone (2×15 mL).The solid material was suspended in EtOH (25 mL). The suspension wasstirred at reflux for 1 hour. The mixture was cooled to roomtemperature, the solid material was collected by filtration, washed withacetone (2×15 mL) and dried in a vacuum oven (50° C.), affording thetitle compound, 3.97 g (90%). ¹H NMR (D₂O, 500 MHz) δ ppm 5.71 (m, 1H),5.04 (m, 2H), 3.05 (m, 2H), 2.85 (m, 1H), 2.53 (m, 1H), 2.20 (m, 1H),1.90 (m, 2H), 1.20 (s. 9H). ¹³C (D₂O, 125 MHz) δ ppm 132.26, 116.10,55.15, 55.02, 37.15, 31.97, 24.08, 22.82. ES-MS 234 (M−1).

Preparation of 4-amino-1-phenyl-2-butanesulfonic acid (Compound QQ)

To a −78° C. solution of 1,3-propane sultone (1 eq) in anhydroustetrahydrofuran (THF, 1.8M) was added butyl lithium (2.5 M in hexanes,1.5 eq). The solution was stirred at −78° C. for 30 minutes beforebenzyl bromide (1 eq diluted with THF) was added via syringe pump over a30 minute period. The reaction mixture was stirred at −78° C. for 2hours. The reaction mixture was warmed up to 0° C. before water (100 mL)was slowly added. The organic layer was recovered and diluted with EtOAc(20 mL). The solution was dried with Na₂SO₄, filtered and evaporatedunder reduced pressure. The product was purified on a silica gel pad(90% Hex/EtOAc to 60% Hex/EtOAc) affording the corresponding1-Benzyl-1,3-propane sultone.

To 0° C. ammonium hydroxide (28-30% NH₃, 50 eq) was added via syringepump over a 4 hours period a solution of 1-Benzyl-1,3-propane sultone (1eq) in tetrahydrofuran (0.8 M). The solution was stirred at 0° C. for anadditional 30 minutes. The solvent was co-evaporated with EtOH. Thesolid was suspended in EtOH. The mixture was stirred at reflux for 1hour. After cooling to room temperature, the solid material wascollected by filtration, washed with EtOH (2×20 mL) and dried in avacuum oven (50° C.), affording the title compound (57%). ¹H NMR (500MHz, D₂O) δ (ppm) 1.77 (m, 1H), 1.89 (m, 1H), 2.57 (t, 1H, J=10.5 Hz),2.76 (m, 1H), 2.94 (m, 1H), 3.05 (m, 1H), 3.23 (dd, 1H, J=4.1 Hz, 13.9Hz), 7.22 (m, 5H); ¹³C NMR (125 MHz, D₂O) δ(ppm) 26.89, 36.05, 37.79,59.42, 127.12, 129.03, 129.48, 138.16; ES-MS 230 (M+H).

Preparation of 1-aminohex-5-ene-3-sulfonic acid (Compound QR)

To a −78° C. solution of 1,3-propane sultone (5.0 g, 41 mmol) inanhydrous tetrahydrofuran (THF, 150 mL) was added butyl lithium (2.5 Min hexanes, 18 mL, 45 mmol). The solution was stirred at −78° C. for 30minutes before Allyl bromide (3.5 mL, 41 mmol) was added via syringepump over a 30 minute period. The reaction mixture was stirred at −48°C. for 2 hours. The reaction mixture was warmed up to 0° C. before water(100 mL) was slowly added. The organic layer was recovered and dilutedwith EtOAc (20 mL). The solution was dried with Na₂SO₄, filtered andevaporated under reduced pressure. The product was purified on a silicagel column (100% Hexanes to 80% Hex/EtOAc) affording the corresponding1-allyl-1,3-propane sultone (5.76 g, 43%).

To 0° C. ammonium hydroxide (28-30% NH₃, 230 mL, 1.8 mol), a solution of1-allyl-1,3-propane sultone (6.66 g, 0.041 mol) in tetrahydrofuran (25mL) was added via syringe pump over a 4 hour period. The solution wasstirred at 0° C. for an additional 30 minutes. The solvent wasco-evaporated with EtOH. The solid was suspended in acetone, collectedby filtration and dried in a vacuum oven (50° C.), affording the titlecompound, 4.96 g (68%). ¹H NMR. (D₂O, 500 MHz) δ ppm 5.73 (m, 1H), 5.01(m, 2H), 3.06 (m, 2H), 2.85 (m, 1H), 2.54 (m, 1H), 2.20 (m, 1H), 1.93(m, 2H). ¹³C (D₂O, 125 MHz) δ ppm 134.42, 118.26, 57.24, 37.63, 34.20,26.95. ES-MS 180 (M+1).

Preparation of (R)-(−)-3-(1-methylpropylamino)-1-propanesulfonic acid(Compound QS)

A solution of 1,3-propane sultone (9.35 g, 75 mmol, Avocado A11923 lotD14N12) in toluene (50 mL, Fisher T290-4, lot 041983) was added to asolution of (R)-(−)-2-butylamine (5.45 g, 74 mmol, Lancaster 3889 lotFA018393) in acetone (25 mL, EMD AX0115-1, lot 44215432). The mixturewas heated to reflux for 24 hours. The mixture was cooled to 0° C., andthe solid was collected by suction filtration, rinsed with acetone (2×10mL) and dried under vacuum (13.28 g). The solid was suspended in ethanol(60 mL, ADSQ-7, lot 5730) and the suspension was heated to reflux. Water(0.1 mL) was then added to afford a clear solution. The mixture wasslowly cooled to 0° C. and the solid was collected by suctionfiltration, rinsed with ethanol (2×10 mL) and dried 20 hours at 60° C.in the vacuum oven. The title compound was obtained as a fine whitesolid (10.51 g, 53.82 mmol, 73%). ¹H NMR (300 MHz, D₂O) δ 0.92 (t, J=7.6Hz, 3H), 1.28 (d, J=6.3 Hz, 3H), 1.48-1.63 (m, 1H), 1.71-1.85 (m, 1H),2.10 (q, J=7.7 Hz, 2H), 3.00 (t, J=7.3 Hz, 2H), 3.11-3.29 (m, 3H); ¹³CNMR (75 MHz, D₂O) δ 9.2, 15.3, 21.9, 26.0, 43.6, 48.2, 56.2; ES-MS 194(M−H); [α]_(D)=−1.2±0.1 (c=0.0157, H₂O).

Preparation of (1E,3S)-3-amino-4-phenylbut-1-ene-1-sulfonic acid(Compound QT)

n-BuLi ((24 mL, 60 mmol, 2.5M in THF) was added at −78° C. to a solutionof methanesulfonate (4.17 mL, 40.3 mmol). The mixture was stirred for 15minutes then chlorophosphonate was added dropwise. The reaction wasallowed to slowly warm to room temperature overnight. NH₄Cl solution wasadded and the reaction was extracted with EtOAc. The organic layer waswashed with brine, dried (Na₂SO₄) and concentrated. The crude waspurified by column using Hex:EtOAc 50/50 to obtain 2.5 g (50% yield) ofthe desired product. ¹H NMR (CDCl₃, 500 MHz) δ 1.37 (t, J=7.0 Hz, 6H),1.44 (t, J=7.0 Hz, 3H), 3.71 (d, J=17.0 Hz, 2H), 4.23 (m, 4H), 4.43 (q,J=2H).

Dibal (39 mL, 1M solution in cyclohexane) was added slowly within twohours to a cold (−78° C.) solution of ester (7.2 g, 25.77 mmol) inCH₂Cl₂ (60 mL). After the end of the addition, the reaction was leftstirring at −78° C. for one additional hour before it was quenchedcarefully with MeOH at −78° C. To the resulting white emulsion was addedHCl (1M) (170 mL) and the mixture was stirred for 15 minutes and theaqueous mixture was extracted with CH₂Cl₂. The combined organic layerswere dried over Na₂SO₄, filtered and concentrated. The crude product waspurified by column using Hexanes:EtOAc 70:30 to afford the aldehyde as awhite solid 5 g (78% yield).

To a suspension of NaH (173 mg, 7.23 mmol) in THF (5 mL) was addeddropwise a solution of phosphonate (2.55 g, 9.46 mmol) in THF (40 mL) at0° C. After the end of the addition the reaction was stirred for 15minutes then the aldehyde (1.2 g, 4.82 mmol) was added in one portion.The reaction mixture was stirred at room temperature for 30 minutesbefore being quenched with H₂O and EtOAc. The organic layer was driedover Na₂SO₄, filtered and concentrated. The crude product was purifiedby column using Hex:EtOAc 90:10-70:30 to afford 1.6 g (93%) of thedesired product. ¹H NMR (CDCl₃, 500 MHz) S 1.1.32 (m, 12H0, 3.00 (m,2H), 4.05 (m, 2H), 4.60 (m, 1H), 4.70 (bs, 1H), 6.12 (m, 1H), 6.85 (m,1H), 7.15-7.40 (m, 5H).

A solution of the sulfonate (800 mg, 2.25 mol) in a mixture of formicacid/water (5 mL/0.2 mL) was heated at reflux for 48 hours thenconcentrated under reduced pressure. EtOH (15 mL) was added undervigorous stirring with heating at reflux for 30 minutes. The suspensionwas cooled then diluted with acetone (5 mL) and filtered. The whitesolid was washed with EtOH and Et₂O then dried under high vacuum toobtain 420 mg (82% yield). ¹H NMR (D₂O, 500 MHz) δ 2.94 (d, J=7.3 Hz,2H), 4.10 (m, 1H), 6.33 (m, 2H), 7.10-7.32 (m, 5H). ¹³C (D₂O, 125 MHz) δ38.53, 52.82, 127.77, 129.10, 129.71, 132.70, 134.50. ES-MS 226 (M−1).

Preparation (3S)-3-amino-4-phenylbutane-1-sulfonic acid (Compound QU)

To a stirred solution of the sulfonate (1.4 g, 3.94 mmol) in MeOH wasadded Pd/C (300 mg). The suspension was stirred under H₂ pressure for 3hours then filtered on celite and the filtrate was concentrated toafford a white solid 1.2 g, 86% yield. ¹H NMR (CDCl₃, 500 MHz) δ 1.34(t, J=7 Hz, 3H), 1.39 (s, 9H), 1.83 (m, 1H), 2.10 (m, 1H), 2.80 (m, 2H),3.15 (m, 2H), 3.90 (m, 1H), 4.23 (q, J=7.0 hz, 2H), 4.40 (bd, NH),7.14-7.30 (m, 5H).

A solution of the sulfonate (1.3 g, 3.63 mol) in a mixture of formicacid/water (8 mL/0.4 mL) was heated for 3 days then concentrated underreduced pressure. EtOH (15 mL) was added under vigorous stirring withheating at reflux for 30 minutes. The suspension was cooled then dilutedwith acetone (5 mL) and filtered. The white solid was washed with EtOHand Et₂O then dried under high vacuum to obtain 700 mg (84% yield). ¹HNMR (D₂O, 500 MHz) δ 2.00 (m, 2H), 2.70 and 2.95 (ABX, J=15.0 and 8.0Hz, 2H), 2.88 (m, 2H), 3.55 (m, 1H), 7.14-7.30 (m, 5H). ¹³C (D₂O, 125MHz) δ 27.82, 38.18, 47.29, 52.25, 127.70, 129.22, 129.48, 135.33. ES-MS228 (M−1).

Preparation of 3-{[1-(3-fluorophenyl)propyl]amino}-1-propanesulfonicacid (Compound QV)

Borane solution (120 mL 1M in THF) was added dropwise to a solution ofamino-acid (5 g, 48.54 mmol) in THF (100 mL) and the reaction mixturewas heated at reflux for 15 hours. The reaction was quenched carefullywith MeOH then concentrated under reduced pressure. It was redilutedwith MeOH and concentrated HCl then heated at reflux for one hour andconcentrated under reduced pressure. The residue (˜6 g) was dried underpump vacuum to afford an oil. ¹H NMR (300 MHz, D₂O) δ 0.81 (d, J=7.0 Hz,3H), 1.88 (m, 1H), 2.75 & 2.92 (ABX, J=14.0 & 7.0 Hz, 2H), 3.53 &3.45(ABX, J=14.0 and 7.0 Hz, 2H).

A solution of the crude amino-alcohol (6 g, 48 mmol) in anhydrous CHCl₃was saturated with HCl(g) and then treated dropwise at reflux withSOCl₂. After the addition was completed, precipitation of a white solidoccurred. After refluxing for 1 hour, the reaction mixture became clear.The reaction was concentrated to yield a colorless syrup which did notprecipitate by trying different solvent systems. ¹H, ¹³C NMR and MSshowed a mixture of at least two products including the desiredchloride.

A solution of the crude chloride in water (5 mL) was added dropwise to arefluxed solution of N₂SO₃ (8 g, 63.5 mmol) in water (25 mL). After theend of the addition, the reaction was stirred at reflux for 20 minutesthen cooled down and concentrated under reduced pressure. 16 mL of HCl(cone) were added to dissolve the aminosulfonic acid and precipitate theinorganic salts which were removed by filtration. The filtrate wasconcentrated, then ethanol was added to cause amino sulfonic acid toappear as white solid which was collected by filtration. It was washedwith EtOH and Et₂O, then dried under high vacuum to obtain 1.9 g of awhite solid (25% yield over three steps). ¹H NMR (500 MHz, D₂O) δ 1.05(d, J=7.0 Hz, 3H), 2.27 (m, 1H), 2.84 & 2.88 (ABX J=15.0 and 7.0 Hz,2H), 2.86 & 3.10 (ABX, J=15.0 and 7.0 Hz, 2H). ¹³NMR (125 MHz, D₂O)δ17.26, 28.90, 44.49, 54.99. ES-MS 152 (M−1).

Preparation of 4-(2-aminoethyl)hepta-1,6-diene-4-sulfonic acid (CompoundN2)

To a −48° C. solution of 1,3-propane sultone (5.0 g, 41 mmol) inanhydrous THF (150 mL) was added butyl lithium (2.5 M in hexanes, 18 mL,45 mmol). The solution was stirred at −78° C. for 0.5 hours before allylbromide (3.5 mL, 41 mmol) was added via syringe pump over 0.5 hourperiod. The reaction mixture was stirred at −48° C. for 2 hours. Thereaction mixture was warmed to 0° C. before water (100 mL) was slowlyadded. The organic layer was recovered. The aqueous layer was extractedwith EtOAc (3×100 mL). The organic extracts were combined, dried overNa₂SO₄, filtered and evaporated under reduced pressure. The product waspurified on a silica gel column (90% Hexanes/EtOAc to 70%Hexanes/EtOAc), affording the corresponding 1,1-diallyl-1,3-propanesultone (0.73 g).

To an aqueous solution of ammonium hydroxide (28-30% NH₃, 21 mL, 180mmol) at 0° C. was added a solution of 1,1-diallyl-1,3-propane sultone(0.73 g, 3.6 mmol) in THF (5 mL) via syringe pump over a 4 hour period.The solution was stirred at 0° C. for an additional 0.5 hours. Thesolvent was co-evaporated with EtOH. The residue was dissolved in water(15 mL) before the solution was extracted with EtOAc (3×15 mL). Theaqueous phase was recovered and evaporated to dryness and lyophilized,affording the title compound (493 mg, 63%). ¹H NMR (D₂O, 500 MHz) δ ppm5.81 (m, 2H), 5.04 (m, 4H), 3.08 (m, 2H), 2.37 (t, 4H, J=8.3 Hz), 1.86(m, 2H); ¹³C (D₂O, 125 MHz) δ ppm 133.40, 119.81, 62.21, 38.38, 35.86,31.97; ES-MS 217 (M−1).

Preparation of3-(1,1-dimethyl-2-methoxy-2-oxoethyl)amino-1-propanesulfonic acid(Compound N3)

Methyl α-aminoisobutyrate hydrochloride (10.0 g, 65.1 mmol) was treatedwith a saturated aqueous solution of K₂CO₃ (50 mL) and EtOAc (3×50 mL).The organic extracts were combined, dried over Na₂SO₄, evaporated underreduced pressure and dried in vacuo to give methyl α-aminoisobutyrate.

To a solution of methyl α-aminoisobutyrate (3.58 g, 30.6 mmol, fromstep 1) in a solvent mixture of toluene and acetonitrile (40 mL,v/v=1:3) was added 1,3-propane sultone (3.56 g, 29.1 mmol). The solutionwas stirred at reflux overnight. The reaction mixture was cooled to roomtemperature. The solid was collected by filtration, washed with acetone(2×25 mL) and dried in a vacuum oven (50° C.), affording the titlecompound (6.35 g, 87%); ¹H NMR (D₂O, 500 MHz) δ ppm 3.87 (s, 3H), 3.24(t, 2H, J=7.3 Hz), 3.03 (t, 2H, J=7.3 Hz), 2.17 (m, 2H), 1.62 (s, 6H);¹³C (D₂O, 125 MHz) δ ppm 172.44, 62.64, 54.21, 48.12, 42.16, 22.01,21.47; ES-MS 238 (M−1).

Preparation of 3-amino-2-benzyl-1-propanesulfonic acid (Compound N4)

To a cold (−78° C.) solution of 3-hydroxypropionitrile (1 g, 14.06 mmol)in THF (30 mL), was added a solution of lithium bis(trimethylsilyl)amide(1 M in THF, 28 mL). After the reaction mixture was stirred for 1 h at−78° C., benzyl bromide (1.67 mL, 14.06 mmol) was added dropwise and thereaction mixture was warmed to reach 0° C. at which temperature themixture was stirred overnight. The reaction was quenched with 1N HCl andextracted with EtOAc. The organic layer was washed with 1N HCl, driedover Na₂SO₄ and concentrated. The residue was applied on silica gelcolumn (eluant:Hexanes:EtOAc 70:30 to 50:50) to afford 1.3 g (69%) ofthe 2-benzyl-3-hydroxypropionitrile. ¹H NMR (300 MHz, CDCl₃) δ 2.80 (bs,1H), 2.95 (m, 3H), 3.77 (m, 2H), 7.20-7.35 (m, 5H); ¹³C NMR (125 MHz,CDCl₃) δ 34.71, 37.03, 61.98, 120.78, 127.58, 129.06, 129.25, 136.71.The dialkylated product was isolated in 8.5% yield.

To a solution of 2-benzyl-3-hydroxypropionitrile (obtained in step 1, 3g, 24.75 mmol) in EtOH (60 mL) was added an aqueous solution of NH₄OH(30%, 20 mL), followed by Ra-Ni (3 g). The suspension was stirred underatmosphere H₂ pressure for 15 hours and then filtered. The filtrate wasconcentrated under high vacuum; and the residual product(3-1mino-2-benzyl-1-propanol) was used in the next step withoutpurification.

A solution of the crude 3-amino-2-benzyl-1-propanol (4.5 g, 27.23 mmol)in anhydrous CHCl₃ (24 mL) was saturated with HCl (g), and then SOCl₂(5.2 mL, 71.0 mmol) was added dropwise at reflux. The reaction wasmaintained under reflux for an additional 2 hours. The reaction was thenconcentrated to yield a syrupy product. The crude3-chloro-2-benzyl-1-propylamine thus obtained was used in the next stepwithout further purification.

A solution of the crude 3-chloro-2-benzyl-1-propylamine (obtained instep 3) in water (10 mL) was added dropwise to a solution of Na₂SO₃ (6.8g, 54.46 mmol) in water (25 mL) under reflux. After the end of theaddition, the reaction was stirred at reflux for 1 hour, then cooleddown and concentrated under reduced pressure. HCl (conc. 16 mL) wereadded to dissolve the aminosulfonic acid and precipitate the inorganicsalts which were removed by filtration. The filtrate was concentrated;and ethanol was added. The title amino sulfonic acid was precipitated aswhite solid which was collected by filtration, washed with EtOH andEt₂O, then dried under high vacuum to give a white solid (1.87 g, 30%yield over three steps). ¹H NMR (500 MHz, D₂O) δ 2.52 (m, 1H), 2.8 (m,2H), 2.94 (m, 2H), 3.08 & 3.18 (ABX, J=13.0 & 7.0 Hz, 2H), 7.25-7.37 (m,5H). ¹³C NMR (125 MHz, D₂O) δ 35.47, 37.78, 42.67, 52.55, 127.15,129.09, 129.54, 138.32. ES-MS 228 (M−1).

Preparation of 1-aminopentane-3-sulfonic acid (Compound N5)

To a −78° C. solution of 1,3-propane sultone (5.0 g, 41.0 mmol) inanhydrous THF (150 mL) was added butyl lithium (2.5 M in hexanes, 18 mL,45 mmol). The solution was stirred at −78° C. for 0.5 h beforeiodoethane (3.3 mL, 41 mmol) was added via syringe pump over 0.5 hourperiod. The reaction mixture was stirred at −78° C. for 4 hours. Thereaction mixture was warmed up to 0° C. before water (100 mL) was slowlyadded. The organic layer was recovered. The aqueous layer was extractedwith EtOAc (3×100 mL). The organic extracts were combines, dried overNa₂SO₄, and evaporated under reduced pressure. The residual material waspurified on a silica gel column (Hexanes/EtOAc from v/v=9:1 to v/v=8:2),affording the 1-ethyl-1,3-propane sultone (3.19 g, 52%).

To a 0° C. aqueous solution of ammonium hydroxide (28-30%, 153 mL, 1.31mol) was added via syringe pump over a 4 hour period a solution of1-ethyl-1,3-propane sultone (3.19 g, 26.4 mmol) in THF (26 mL). Thesolution was stirred at 0° C. for 3 hours and at room temperatureovernight. The solvent was co-evaporated with EtOH. The solid materialwas treated acetone (150 mL), collected by filtration, and dried in avacuum oven (50° C.), affording the title compound (2.96 g, 67%); ¹H NMR(D₂O, 500 MHz) δ ppm 3.21 (m, 2H), 2.83 (m, 1H), 2.09 (m, 2H), 1.93 (m,1H), 1.63 (m, 1H), 1.03 (t, 3H, J=7.3 Hz); ¹³C (D₂O, 125 MHz) δ ppm59.39, 37.75, 26.90, 22.87, 10.69; ES-MS 166 (M−1).

General Procedure for the Synthesis of Compounds Starting from β-AminoAcids.

A solution of borane:tetrahydrofurane complex (1M, 3-4 mL per mmol=3-11equiv. of β-amino acid) was added dropwise over a period of 1 hour to acold (0° C.) suspension of beta amino acid (1 equiv.) in THF (1 mL permmol). The mixture was stirred 20 minutes at room temperature at the endof the addition. It was then heated at reflux for 22 hours. The mixturewas then cooled to 0° C.; and methanol (2 mL per mmol) was added over aperiod of 30 minutes. The mixture was then heated at reflux for 20minutes and concentrated to a thick oil. The oil was co-evaporated withmethanol (3×200 mL). The solid obtained was dried in vacuo to afford thecorresponding 3-substituted 3-amino-1-propanol derivative as a whitewaxy solid (quantitative yield).

An aqueous solution of HBr (48%, 2 mL per equiv. of the alcohol) wasadded slowly to a flask containing a 3-substituted 3-amino-1-propanol (1equilv.). The mixture was heated at reflux for 6 hours, and thenconcentrated to dryness. The crude material was used directly in thenext step.

The 1-substituted 3-bromo-1-propylamine hydrobromide (obtained in step2) was added to a solution of sodium sulfite (1.0 equiv. of the1-substituted 3-bromo-1-propylamine) in water and 1,4-dioxane. Themixture was heated under reflux for 6 h then concentrated to dryness.The residual material was treated with concentrated HCl. The inorganicmaterial was removed by filtration; and the filtrate was treatedethanol, causing precipitation of the corresponding sulfonic acid. Thecrude sulfonic acid was suspended in ethanol and the mixture was heatedat reflux for 1 hour. After cooling to room temperature, the solidmaterial was collected by filtration, rinsed with ethanol and driedovernight in the vacuum oven at 60° C., giving the corresponding3-substituted 3-amino-1-propanesulfonic acid as a fine white crystallinesolid.

3-amino-1-butanesulfonic acid, (Compound N6)

Overall yield 5.4% (0.29 g); NMR (500 MHz, D₂O) δ 1.22 (d, J=6.8 Hz,3H), 1.85-1.93 (m, 1H), 1.99-2.06 (m, 1H), 2.86-2.96 (m, 2H), 3.04-3.44(m, 1H); ¹³C (125 MHz, D₂O) δ 17.6, 29.5, 46.9, 47.3; ES-MS 152 (M−1)

3-amino-3-cyclohexyl-1-propanesulfonic acid, (Compound N10)

Overall yield 60% (14.5 g); ¹H NMR (500 MHz, D₂O) δ 0.87-1.04 (m, 3H),1.07-1.17 (m, 2H), 1.47-1.65 (m, 6H), 1.84-1.92 (m, 1H), 1.99-2.06 (m,1H), 2.81-2.91 (m, 2H), 3.11-3.15 (m, 1H); ¹³C (125 MHz, D₂O) δ25.0,25.5, 25.6, 25.7, 27.4, 28.2, 39.1, 47.2, 55.5; ES-MS: 220 (M−1)

3-amino-1-heptanesulfonic acid, (Compound N16)

Overall yield 48% (3.1 g); ¹H NMR (500 MHz, D₂O) δ 0.73 (t, J=5.9 Hz,3H), 1.20 (very br s, 4H), 1.46-1.57 (m, 2H), 1.87-1.99 (m, 2H), 2.86(t, J=7.8 Hz, 2H), 3.26-3.31 (m, 1H); ¹³C (125 MHz, D₂O) δ 14.1, 21.8,26.4, 27.4, 31.3, 47.0, 50.8; ES-MS (M−1)

3-amino-5-methyl-1-hexanesulfonic acid, (Compound N17)

¹H NMR (500 MHz, D₂O) δ 0.76-0.77 (m, 6H), 1.37 (oct, J=7.4 Hz, 2H),1.54 (hep, J=6.8 Hz, 1H), 1.92 (dec, J=7.3 Hz, 2H), 2.84-2.90 (m, 2H),3.56 (qt, J=6.7 Hz, 1H); ¹³C (125 MHz, D₂O) δ 21.3, 31.8, 23.9, 27.9,41.0, 46.9, 49.1; ES-MS 194 (M−1)

Preparation of 3-cycloheptylmethyl-1-propansulfonic acid (Compound N7)

A solution of 1,3-propane sultone (1.20 g, 9.5 mmol) in toluene (6 mL)was added to a solution of cycloheptanemethylamine (1.19 g, 9.35 mmol)in acetone (6 mL). The mixture was stirred at reflux for 4 hours.Ethanol (10 mL) was added and the mixture was cooled to roomtemperature. The solid was collected by suction-filtration, rinsed withethanol (2×5 mL), and dried overnight at 60° C. in the vacuum oven,giving the title compound as a white solid (1.83 g, 79%); ¹H NMR (500MHz, DMSO-d6) δ 1.18-1.20 (m, 2H), 1.41-1.79 (m, 11H), 1.95 (br s, 2H),2.64 (br s, 2H), 2.74-2.76 (m, 2H), 3.04 (br s, 2H), 5.51 (br s, 2H);¹³C (125 MHz, DMSO-d6) δ 21.6, 25.3, 27.8, 31.1, 36.1, 47.6, 49.4, 53.0;ES-MS 248 (M−1)

Preparation of3-[(R)-(3-benzylthio-1-hydroxy-2-propyl)amino]-1-propansulfonic acid(Compound N8)

A solution of S-benzyl-L-cysteol (1 g, 4.9 mmol) in acetone (6 mL) wasfiltered on paper. To the solution was added a solution of 1,3-propanesultone (0.70 g, 5.5 mmol) in toluene (6 mL). The mixture was stirred atreflux for 4 hours. Ethanol (5 mL) was added and the mixture was cooledto room temperature. The solid was collected by filtration, rinsed withethanol (2×2.5 mL) then dried overnight at 60° C. in a vacuum oven,giving the title compound as a white solid (0.85 g, 54%); ¹H NMR (500MHz, DMSO-d6) δ 1.94 (qt, J=6.7 Hz, 2H), 2.59-2.74 (m, 4H), 3.08 (m,2H), 3.22 (m, 1H), 3.34 (s, 1H), 3.82 (t, J=3.7 Hz, 1H), 5.37 (T, J=4.7Hz, 1H), 7.22-7.36 (m, 5H), 8.64 (BR S, 2H); ¹³C (125 MHz, DMSO-d6)521.9, 28.2, 35.3, 44.8, 49.1, 57.3, 57.5, 126.7, 128.2, 128.7, 137.8;ES-MS 318 (M−1); [α]_(D)−13.5±0.1 (c=0.0103 in water).

Preparation of 1-amino-5-methyl-3-hexanesulfonic acid (Compound N9)

To a −78° C. solution of 1,3-propane sultone (10 g, 82 mmol) inanhydrous THF (300 mL) was added butyl lithium (2.5 M in hexanes, 36 mL,90 mmol). The solution was stirred at −78° C. for 0.5 hours before1-iodo2-methylpropane (9.5 mL, 82 mmol) was added via syringe pump over0.5 hour period. The reaction mixture was stirred at −78° C. for 4hours. The reaction mixture was warmed up to 0° C. before water (200 mL)was slowly added. The organic layer was recovered. The aqueous layer wasextracted with EtOAc (3×100 mL). The organic extracts were combined,dried over Na₂SO₄, and evaporated under reduced pressure. The residualmaterial was purified on a silica gel column (100% Hexanes to 80%Hexanes/EtOAc), affording 1-isobutyl-1,3-propane sultone (0.3 g, 2%).

Step 2: To an aqueous solution of ammonium hydroxide (28-30%, 10 mL, 85mmol) at 0° C. was added via syringe pump over a 4-h period a solutionof 1-isobutyl-1,3-propane sultone (0.3 g, 1.7 mmol) in THF (5 mL). Thesolution was stirred at 0° C. for 1 hour and at room temperatureovernight. The solvent was co-evaporated with EtOH. The solid was driedin a vacuum oven (50° C.), affording the title compound (0.258 g,78.7%). ¹H NMR (D₂O, 500 MHz) δ ppm 3.21 (m, 2H), 2.95 (m, 1H), 2.06 (m,2H), 1.71 (m, 2H), 1.45 (m, 1H), 0.94 (d, 3H, J=6.3 Hz), 0.94 (d, 3H,J=6.3 Hz), 0.90 (d, 3H, J=6.3 Hz); ¹³C (D₂O, 125 MHz) δ ppm 56.19,38.69, 37.58, 27.77, 25.19, 22.55, 20.97; ES-MS 196 (M+1).

Preparation of 6-(aminomethyl)-3,4-dimethylcyclohex-3-ene-1-sulfonicacid (Compound N11)

To a cold (−40° C.) solution of allyl alcohol (20 ml, 300 mmol) and NEt₃(26 mL, 186 mmol) in THF (150 mL) was added dropwise2-chloroethanesulfonyl chloride (10.4 mL, 100 mmol). The reaction wasstirred at −40 to −20° C. for 5 hours, quenched with HCl (1M) andextracted with EtOAc. The organic layer was washed with water and driedover Na₂SO₄. The product was purified by column chromatography usingHexanes/EtOAc 80/20 as eluant to afford allyl vinylsulfonate as ayellowish oil (7 g, 47%). ¹H NMR (500 MHz, CDCl₃) δ 4.55 (m, 2H), 5.34(m, 2H), 5.85 (m, 1H), 6.06 (d, J=6.0 Hz, 1H), 6.35 (d, J=17.0 Hz, 1H),6.50 (dd, J=17 & 9.5 Hz, 1H).

To a degassed (by Nitrogen bubbling) solution of allyl vinylsulfonate (3g, 20.24 mmol) in CH₂Cl₂ (1 L) was added Grubbs Catalyst (170 mg, 0.2mmol). The reaction was heated at reflux for 2 h then concentrated. Theresidual material was applied on silica gel column using Hexanes/EtOAc80/20 to 50/50 as eluant to afford 1,3-prop-1-ene sultone 2.2 g (92%).¹H NMR (500 MHz, CDCl₃) δ 5.11 (dd, J=2.2 & 2.2 Hz, 2H), 6.80 (dt, J=6.6& 2.2 Hz, 1H), 7.00 (dt, J=6.6 & 2.0 Hz, 1H); ¹³C NMR (125 MHz, CDCl₃) δ72.54, 124.76, 137.04.

A mixture of 1,3-propene sultone (1.44 g, 12 mmol),2,3-dimethyl-1,3-butanediene (9.5 mL, 84 mmol) in 30 mL of toluene wasplaced in a sealed tube and heated at 150° C. for 15 hours. The solventwas removed and the residual material was applied on silica gel columnusing Hexanes/EtOAc 80:20 to 70:30 as eluant to afford 700 mg (86%) ofthe Diels-Alder adduct. ¹H NMR (500 MHz, CDCl₃) δ 1.62 (s, 3H), 1.66 (s,3H), 1.88-1.92 (m, 1H), 2.28-2.42 (m, 3H), 3.12-3.18 (m, 1H), 3.48 (q,J=7.6 Hz, 1H), 3.96 (t, J=8.5 Hz, 1H), 4.40 (dd, J=8.50 & 7.25 Hz, 1H).¹³C NMR (125 MHz, CDCl₃) δ 19.13, 19.25, 27.20, 30.79, 34.18, 53.04,72.61, 122.70, 123.44.

To an ice-cooled solution of NH₄OH (28% in water, 22 mL, 168 mmol) in aco-solvent of THF and EtOH (20 mL, v/v=1:1) was added slowly via asyringe pump a solution of the Diels-Alder adduct from step 3 (680 mg,3.36 mmol). After the addition (2 h), the reaction was stirred for twomore hours until TLC indicated complete consumption of the startingmaterial. The solvent was evaporated and the resulting solid wassuspended in mixed solvents of EtOH, acetone and ether, heated for 15minutes and cooled. The solid was collected by filtration, washed withether and dried, to give the title compound (450 mg, 61%). ¹H NMR (500MHz, D₂O) δ 1.49. (s, 3H), 1.52 (s, 3H), 1.86-1.92 (m, 1H), 2.14-2.28(m, 3H), 2.44-2.49 (m, 1H), 2.80 (dd, J=13.0 & 6.0 Hz, 1H), 3.10-3.16(m, 1H), 3.27 (dd, J=14.0 & 6.0 Hz, 1H); ¹³C NMR (125 MHz, D₂O) δ 18.02,18.28, 28.93, 32.36, 34.69, 38.79, 57.85, 123.09, 123.72. ES-MS 218(M−1).

Preparation of6-[(tert-butylamino)methyl]-3,4-dimethylcyclohex-3-ene-1-sulfonic acid(Compound N13)

To a solution of the Diels-Alder adduct from step 3 in the synthesis ofCompound N11 (607 mg, 3 mmol) in pinacolone (6 mL) was addedt-butylamine (381 μL, 3.6 mmol). The mixture was stirred at reflux for 3hours then another 381 μL of amine was added. The reaction was stirredat reflux for 2 more hours then concentrated. The solid was suspended inEtOH and heated for 15 minutes and then cooled. The solid was collectedby filtration, giving the title compound (720 mg, 87%); ¹H NMR (500 MHz,DMSO-d₆) δ 1.22 (s, 9H), 1.57 (s, 3H), 1.60 (s, 3H), 1.84-1.90 (m, 1H),2.03 (m, 2H), 2.36-2.42 (m, 1H), 2.58 (m, 1H), 2.64 (m, 1H), 2.82 (m,1H), 3.00 (m, 1H); ¹³C NMR (125 MHz, DMSO-d₆) δ 19.33, 19.48, 25.81,29.67, 31.78, 38.40, 41.38, 56.10, 57.64, 123.30, 124.02. ES-MS 274(M−1).

Preparation of6-(2-adamantylamino)methyl-3,4-dimethylcyclohex-3-ene-1-sulfonic acid(Compound N14)

To a solution of the Diels-Alder adduct from step 3 in the synthesis ofCompound N11 (607 mg, 3 mmol) in pinacolone (6 mL) was added2-adamantaneamine (544 mg, 3.6 mmol). The reaction was stirred at refluxfor 5 hours then concentrated. The solid was suspended in MeOH andheated for 15 min and then cooled. The solid was collected byfiltration, affording the title compound (260 mg, 24%). ¹H NMR (500 MHz,DMSO-d₆) δ 1.25-1.69 (m, 9H), 1.70 (m, 5H), 1.80 (s, 3H), 1.64 (s, 3H),2.00 (m, 1H), 2.08 (m, 2H), 2.23 (m, 2H), 2.41 (m, 1H), 2.50 (s, 1H),2.73 (m, 2H), 2.82 (m, 1H), 3.02 (m, 1H), 3.20 (m, 1H), 8.49 (bs, 1H),9.31 (bs, 1H); ¹³C NMR (125 MHz, DMSO-d₆) δ 19.30, 19.46, 26.82, 27.12,28.15, 29.64, 30.03, 30.25, 30.47, 31.59, 36.69, 36.89, 37.29, 38.89,46.14, 57.43, 62.73, 123.23, 124.04. ES-MS 352 (M−1).

Preparation of 1-amino-4-hydroxy-4-methyl-3-pentanesulfonic acid(Compound N15)

To a −78° C. solution of 1,3-propane sultone (5.0 g, 41 mmol) inanhydrous THF (150 mL) was added butyl lithium (2.5 M in hexanes, 18 mL,45 mmol). The solution was stirred at −78° C. for 0.5 hours beforeacetone (3.0 mL, 41 mmol) was added via syringe pump over a 0.5-hperiod. The reaction mixture was stirred at −78° C. for 4 hours. Themixture was warmed up to 0° C. before water (100 mL) was slowly added.The organic layer was separated. The aqueous layer was extracted withEtOAc (3×100 mL) and combined with the organic layer. The combinedorganic extracts were dried over Na₂SO₄ and evaporated under reducedpressure. The residue was separated on a silica gel column (100% Hexanesto 80% Hexanes/EtOAc), affording the corresponding sultone derivative(4.36 g, 60%).

To an aqueous solution of ammonium hydroxide (28-30%, 140 mL, 12.1 mol)at 0° C. was added via syringe pump over a 4 hour period a solution ofsultone derivative from step 1 (4.36 g, 24.2 mmol) in THF (25 mL). Themixture was stirred at 0° C. for 1 hour and at room temperatureovernight. The solvent was co-evaporated with EtOH. The solid was driedin a vacuum oven (50° C.), affording the title compound (4.37 g, 92%).¹H NMR (D₂O, 500 MHz) δ ppm 3.10 (m, 1H), 3.05 (m, 1H), 2.80 (m, 1H),1.26 (d, 3H, J=8.8 Hz), 1.19 (d, 3H, J=8.8 Hz); ¹³C (D₂O, 125 MHz) δ ppm72.26, 67.22, 39.38, 26.80, 26.10, 25.55; ES-MS 198 (M+1).

Preparation of 3-(2-benzylthio-1-ethylamino)-1-propansulfonic acid(Compound N18)

S-Benzylcystamine hydrochloride (2.40 g, 11.8 mmol) was dissolved inwater and the pH was adjusted to basic with potassium carbonate. Theaqueous mixture was extracted with ethyl acetate (3×15 mL). The combinedorganic layers were washed with brine, dried over anhydrous sodiumsulfate; and the solvent was removed under reduced pressure. Theresidue, a mixture of yellowish oil and white solid, was taken inacetonitrile (10 mL); and the mixture was filtered through filter paperand the residue on the paper was rinsed with toluene (10 mL). To thehomogenous organic solution was added 1,3-propane sultone (1.00 mL,containing 11 mmol 1,3-propane sultone). The mixture was stirred atreflux for 20 hours, and then cooled to room temperature. The solid wascollected by suction-filtration, rinsed with acetone (2×5 mL) then driedunder vacuum for 1 hour. The solid was suspended in ethanol (10 mL) andthe mixture was heated under reflux for 1 hour and then cooled to roomtemperature. The solid was collected by suction-filtration, washed withethanol (2×3 mL), dried at 60° C. for 4 hours, providing the titlecompound as a white solid (1.60, 47%); ¹H NMR (500 MHz, DMSO-d6) δ 1.93(qt, J=6.7 Hz, 2H), 2.60-2.64 (m, 4H), 3.08 (v br d, 4H), 3.81 (s, 2H),7.26 (t, J=7.6 Hz, 1H), 7.32-7.39 (m, 4H), 8.62 (br s, 2H); ¹³C (125MHz, DMSO-d6) δ 21.7, 26.2, 34.5, 45.4, 46.7, 48.9, 127.0, 128.5, 129.0,138.1; ES-MS 288 (M−1).

Preparation of6-(1-adamantylamino)methyl-3,4-dimethylcyclohex-3-ene-1-sulfonic acid(Compound N19)

To a solution of the Diels-Alder adduct from step 3 in the preparationof Compound N11 (607 mg, 3 mmol) in pinacolone (6 mL) was added1-adamantaneamine (544 mg, 3.6 mmol). The reaction mixture was stirredat reflux for 5 hours, and then concentrated to dryness. The solidresidue was suspended in MeOH, heated for 15 minutes, cooled to roomtemperature, and collected by filtration, giving the title compound (260mg, 24%); ¹H NMR (500 MHz, DMSO-d₆) δ 1.54-1.66 (m, 11H), 1.75 (s, 3H),1.77 (s, 3H), 1.84 (m, 1H), 2.10 (s, 4H), 2.22 (m, 2H), 2.40 (m, 1H),2.58 (m, 1H), 2.64 (m, 1H), 2.84 (m, 1H), 3.00 (m, 1H), 8.30 (bs, 1H),9.40 (bs, 1H); ¹³C NMR (125 MHz, DMSO-d₆) δ 19.34, 19.48, 29.05, 29.36,31.63, 35.84, 38.42, 38.55, 39.54, 56.23, 57.71, 123.33, 124.01. ES-MS352 (M−1).

Preparation of (Z)-3-(tert-butylamino)prop-1-ene-1-sulfonic acid(Compound N20)

To a boiling solution of allyl bromide (21.6 mL, 250 mmol) in a solventmixture of EtOH and H₂O (200 mL, v/v=3:1) was added dropwise a solutionof sodium sulfite (15.75 g, 125 mmol) in water (60 mL). The reactionmixture was heated under reflux for 3 hours, and concentrated to drynessunder reduced pressure. The obtained white solid was suspended in EtOHin water (130 mL, 90%), heated for 30 minutes, cooled to roomtemperature, and collected by filtration, giving sodiumprop-2-ene-1-sulfonate (14 g, 76%); ¹H NMR (500 MHz, D₂O) δ 3.55 (d,J=7.3 Hz, 2H), 5.35-5.41 (m, 2H), 5.85-6.00 (m, 1H).

To a stirred solution of sulfonate obtained from step 1 (12.0 g, 84mmol) in water (48 mL) was added bromine (about 4.5 mL) dropwise withstirring until the solution turned pale brown. The solution was stirredat room temperature for 3 hours. A small amount of Na₂SO₃ was added todestroy the excess bromine. The solvent was then removed in vacuo and awhite solid was obtained. Without further purification, the2,3-dibromo-1-propanesulfonate was treated with concentrated HCl (50 mL)by stirring at room temperature for 1 day. The precipitate (inorganicsalt) was removed by filtration. The filtrate was concentrated to yellowsyrup. Without further purification, the syrup residue was subjected tovacuum distillation at 140-150° C. to give 2-bromo-1,3-propane sultone(6.5 g, 32%); ¹H NMR (500 MHz, CDCl₃) 8 3.52 (dd, J=14.0 & 7.0 Hz, 1H),3.88 (dd, J=14.0 & 7.0 Hz, 1H), 4.50-4.60 (m, 1H), 4.70-4.82 (m, 2H).

To a solution of 2-bromo-1,3-propane sultone (obtained in Step 2, 8.0 g,39.80 mmol) in toluene (200 mL) was added NEt₃ (9 mL, 65 mmol). Thereaction mixture was stirred for 3 h (or until complete consumption ofthe starting material), diluted with an aqueous solution of HCl (1 M),and extracted twice with EtOAc. The organic layer was dried over Na₂SO₄and concentrated to give 1,3-prop-1-ene sultone (4.5 g, 94%) as a whitesolid; ¹H NMR (500 MHz, CDCl₃) δ 5.11 (dd, J=2.2 & 2.2 Hz, 2H), 6.80(dt, J=6.6 & 2.2 Hz, 1H), 7.00 (dt, J=6.6 & 2.0 Hz, 1H); ¹³C NMR (125MHz, CDCl₃) δ 72.54, 124.76, 137.04.

To a solution of 1,3-prop-1-ene sultone (obtained in step 3, 36 mg, 3mmol) in THF (5 mL) was added tert-butylamine (316 μL, 3 mmol). Thereaction mixture was refluxed for 4 h, and then concentrated to dryness.The residual solid material was suspended in a solvent mixture of EtOH,acetone and ether, heated for 15 minutes, and cooled to roomtemperature. The solid was collected by filtration, washed with etherthen dried, providing the title compound (130 mg, 22%); ¹H NMR (500 MHz,D₂O) δ 1.24 (s, 9H), 4.00 (d, J=7.0 Hz, 2H), 5.94 (m, 1H), 6.50 (d,J=11.0 Hz, 1H). ¹³C NMR (125 MHz, D₂O) δ 25.08, 37.84, 57.63, 127.81,136.08. ES-MS 192 (M−1).

Preparation of (1Z)-(3-(1-adamantylamino)prop-1-ene-1-sulfonic acid(Compound N21)

To a solution of 1,3-prop-1-ene sultone (obtained in step 3 in thepreparation of Compound N20, 360 mg, 3 mmol) in THF (5 mL) was added1-adamantylamine (545 mg, 3.6 mmol). The reaction mixture was refluxedfor 6 h, and then concentrated to dryness. The residual solid wassuspended in a solvent mixture of EtOH and ether (2.5 mL/2.5 mL), heatedunder reflux for 15 minutes, and cooled to room temperature. The solidmaterial was collected by filtration, washed with ether, and dried;providing the title compound (130 mg, 22%); ¹H NMR (500 MHz, D₂O) δ1.40-1.70 (m, 12H), 2.00 (m, 3H), 3.7 (d, J=7.0 Hz, 2H), 5.90 (m, 1H),6.30 (d, J=11.0 Hz, 1H). ¹³C NMR (125 MHz, D₂O) δ 28.95, 29.23, 35.05,35.68, 36.67, 40.05, 40.75, 128.00, 132.98. ES-MS 270 (M−1).

Preparation of (1Z)-3-aminoprop-1-ene-1-sulfonic acid (Compound N22)

A solution of 1,3-prop-1-ene sultone (obtained in step 3 for thepreparation Compound N20, 360 mg, 3 mmol) in a solvent mixture of THFand EtOH (10 mL, v/v=1:1) was added slowly to an aqueous solution ofammonium hydroxide (28%, 26 mL, 200 mmol). The reaction mixture wasstirred at room temperature for 4 hours and then concentrated todryness. The resulting solid was suspended in EtOH, heated at reflux for15 min, and cooled. The solid was collected by filtration, washed withether, and dried, giving the title compound (500 mg, 91%); ¹H NMR (500MHz, D₂O) δ 3.95 (d, J=6.3 Hz, 2H), 5.96 (m, 1H), 6.44 (d, J=11.5 Hz,1H). ¹³C NMR (125 MHz, D₂O) δ 36.25, 129.10, 135.10. ES-MS 136 (M−1).

Preparation of anti-4-amino-1-hydroxy-1-phenyl-2-butanesulfonic acid,trifluoroacetic acid salt (Compound N23),syn-4-amino-1-hydroxy-1-phenyl-2-butanesulfonic acid, trifluoroaceticacid salt (Compound N24) and 4-amino-1-hydroxy-1-phenyl-2-butanesulfonicacid, trifluoroacetic acid salt (Compound N25)

To a −78° C. solution of 1,3-propane sultone (5.0 g, 41 mmol) inanhydrous THF (150 mL) was added butyl lithium (2.5 M in hexanes, 18 mL,45 mmol). The mixture was stirred at −78° C. for 0.5 hours beforebenzaldehyde (4.2 mL, 41 mmol) was added via syringe pump over 0.5 hourperiod. The reaction mixture was stirred at −78° C. for 4 hours, andthen warmed up to 0° C. before water (100 mL) was slowly added. Theorganic layer was separated; and the aqueous layer was extracted withEtOAc (3×100 mL). The organic phase and extracts were combined, driedover Na₂SO₄, and evaporated under reduced pressure. The residue wasseparated on a silica gel column (100% Hexanes to 70% Hex/EtOAc),affording the corresponding sultone derivative (3.04 g, 33%).

To an aqueous solution of ammonium hydroxide (28-30%, 78 mL, 665 mmol)at 0° C. was added via syringe pump, over a 4 hour period, a solution ofthe sultone derivative (obtained in step 1, 3.04 g, 13.3 mmol) in THF(15 mL). The mixture was stirred at 0° C. for 1 hour and then at roomtemperature overnight. The solvent was co-evaporated with EtOH. Themixture of diastereoisomers was separated by preparative HPLC; and thecorresponding fractions were lyophilized, affording the following threecompounds: anti-4-amino-1-hydroxy-1-phenyl-2-butanesulfonic acid,trifluoroacetic acid salt (Compound N23): mixture of enantiomers (204mg). ¹H NMR (D₂O, 500 MHz) δ ppm 7.30 (m, 5H), 4.87 (d, 1H, J=8.0 Hz),3.17 (m, 1H), 2.86 (m, 1H), 2.66 (m, 1H), 1.74 (m, 1H), 1.65 (m, 1H);¹³C (D₂O, 125 MHz) δ ppm 139.51, 128.93, 127.65, 73.82, 63.14, 37.96,25.40; ES-MS 244 (M−1).

anti-4-amino-1-hydroxy-1-phenyl-2-butanesulfonic acid, trifluoroaceticacid salt (Compound N24)

mixture of enantiomers (131 mg); ¹H NMR (D₂O, 500 MHz) δ ppm 7.30 (m,4H), 7.26 (m, 1H), 5.30 (d, 1H, J=2.8 Hz), 3.05 (m, 1H), 2.94 (m, 1H),2.66 (m, 1H), 1.96 (m, 1H); ¹³C (D₂O, 125 MHz) δ ppm 141.35, 128.84,128.05, 125.91, 71.54, 63.65, 38.44, 22.22; ¹⁹F NMR δ ppm −76.31; ES-MS244 (M−1).

4-amino-1-hydroxy-1-phenyl-2-butanesulfonic acid, trifluoroacetic acidsalt (Compound N25)

mixture of diastereoisomers (165 mg). ¹H NMR (D₂O, 500 MHz) δ ppm 7.30(m, 5H), 5.30 (d, 0.5H, J=2.8 Hz), 4.87 (d, 0.5H, J=8.0 Hz), 3.17 (m,0.5H), 3.05 (m, 0.5H), 2.94 (m, 0.5H), 2.86 (m, 0.5H), 2.66 (m, 1H),1.96 (m, 1H), 1.74 (m, 0.5H), 1.65 (m, 0.5H); ¹³C (D₂O, 125 MHz) δ ppm141.35, 128.84, 128.05, 125.91, 71.54, 63.65, 38.44, 22.22; ES-MS 244(M−1).

Preparation of 3-amino-1-phenyl-1-butanesulfonic acid, trifluoroaceticacid salt: Compound N26)

To a 0° C. solution of α-toluenesulfonyl chloride (5 g, 26 mmol) inanhydrous dichloromethane (100 mL) was added ethanol (3 mL, 52 mmol) andtriethylamine (5.5 mL, 39 mmol). The reaction mixture was stirred at 0°C. for 1 h before water was added (100 mL). The organic layer wasseparated and extracted with 2N HCl (1×100 mL) and Brine (1×100 mL). Theorganic phase was dried over Na₂SO₄, evaporated under reduced pressure,and dried in vacuo, affording ethyl phenylmethanesulfonate (4.76 g,91%).

To a −78° C. solution of ethyl phenylmethanesulfonate (4.76 g, 23.8mmol) in anhydrous THF (100 mL) was added butyl lithium (2.5 M inhexanes, 10 mL, 25 mmol). The mixture was stirred at −78° C. for 1 hbefore allyl bromide (3.1 mL, 35.7 mmol) was added via syringe pump overa 0.5 hour period. The reaction mixture was stirred at −78° C. for 5 h,and then warmed up to 0° C. before water (60 mL) was slowly added. Theorganic layer was separated; and the aqueous layer was extracted withEtOAc (3×60 mL). The combined organic layer and extracts were dried overNa₂SO₄, and evaporated to dryness under reduced pressure, affording thecorresponding sultone (4.83 g, 80%).

To an aqueous solution of ammonium hydroxide (28-30%, 87 mL, 750 mmol)at 0° C. was added a solution of sultone (3.19 g, 15.0 mmol) in THF (25mL) and EtOH (20 mL). The solution was stirred at 70° C. overnight. Thesolvent was co-evaporated with EtOH. The solid residue was purified bypreparative HPLC and the corresponding fractions were combined andlyophilized, affording the title compound in a mixture ofdiastereoisomers (170 mg). ¹H NMR (D₂O, 500 MHz) δ ppm 7.28 (m, 5H),4.02 (m, 0.8H), 3.92 (m, 0.2H), 3.52 (m, 0.2H), 3.27 (m, 0.3H), 2.94 (m,0.5H), 2.44 (m, 1H), 2.21 (m, 1H), 1.12 (d, 1.5H, J=6.3 Hz), 1.08 (d,0.9H, J=6.3 Hz), 1.00 (d, 0.6H, J=6.3 Hz); ¹³C (D₂O, 125 MHz) δ ppm135.15, 133.87, 129.37, 129.35, 129.09, 129.00, 128.91, 128.81, 128.79,128.40, 65.84, 63.60, 63.02, 62.66, 46.67, 45.61, 39.18, 35.84, 34.85,21.15, 18.56, 16.64; ES-MS 228 (M−1).

Preparation of 4-amino-4-methyl-1-phenyl-1-pentanesulfonic acid,trifluoroacetic acid salt: (Compound N27)

To a 0° C. solution of α-toluenesulfonyl chloride (10 g, 52 mmol) inanhydrous dichloromethane (200 mL) was added ethanol (6 mL, 104 mmol)and triethylamine (11 mL, 78 mmol). The reaction mixture was stirred at0° C. for 1 hour before water (200 mL) was added. The organic layer wasseparated and extracted with 2N HCl (1×200 mL) and brine (1×200 mL). Theorganic phase was dried over Na₂SO₄, evaporated under reduced pressure,and dried in vacuo, affording the corresponding ethylphenylmethanesulfonate (9.79 g, 93%).

To a −78° C. solution of ethyl phenylmethanesulfonate (4.59 g, 22.9mmol) in anhydrous THF (100 mL) was added butyl lithium (2.5 M inhexanes, 10 mL, 25 mmol). The mixture was stirred at −78° C. for 1 hourbefore 1-bromo-3-methyl-2-butene (2.9 mL, 25.0 mmol) was added viasyringe pump over a 0.5 hour period. The reaction mixture was stirred at−78° C. for 4 hour, and then was warmed up to 0° C. before water (60 mL)was slowly added. The organic layer was separated. The aqueous layer wasextracted with EtOAc (3×60 mL). The organic layer and extracts werecombined, dried over Na₂SO₄, and evaporated under reduced pressure,affording ethyl 4-methyl-1-phenylpent-3-ene-1-sulfonate (3.93 g, 64%).

A solution of ethyl 4-methyl-1-phenylpent-3-ene-1-sulfonate (3.01 g,11.2 mmol) in 2% TFA/CH₂Cl₂ (20 mL) was stirred at reflux for 24 hours.The solvent was evaporated under reduced pressure. The crude oil wasdissolved in 20% TFA/CH₂Cl₂ (20 mL). The solution was stirred at refluxfor 24 hours. After cooling to room temperature, water (20 mL) wasadded. The organic phase was separated and extracted with a saturatedsolution of NaHCO₃ (1×20 mL). The organic phase was dried over Na₂SO₄,evaporated, and dried in vacuo, affording the corresponding sultonederivative (2.42 g, 90%).

To an aqueous solution of ammonium hydroxide (28-30%, 78 mL, 665 mmol)at 0° C. to ammonium hydroxide (28-30% NH₃, 30 mL) was added a solutionof the sultone derivative obtained from step 3 (2.42 g, 10.1 mmol) inTHF (25 mL) and EtOH (20 mL). The solution was stirred at 70° C.overnight. The solvent was co-evaporated with EtOH. The residual solidwas purified by preparative HPLC and the corresponding fractions werecombined and lyophilized, affording the title compound (222 mg). ¹H NMR(D₂O, 500 MHz) δ ppm 7.27 (m, 5H), 3.82 (dd, 1H, J=3.2 Hz, 11.5 Hz),2.14 (m, 1H), 1.98 (m, 1H), 1.32 (m, 1H), 1.10 (m, 1H), 1.02 (s, 6H);¹³C (D₂O, 125 MHz) δ ppm 135.70, 129.50, 128.92, 128.77, 128.34, 71.34,40.13, 27.74, 27.54, 25.24; ES-MS 256 (M−1).

Preparation of 3-amino-1-phenyl-1-pentanesulfonic acid, trifluoroaceticacid salt: (Compound N30)

To a 0° C. solution of α-toluenesulfonyl chloride (10 g, 52 mmol) inanhydrous dichloromethane (200 mL) was added ethanol (6 mL, 104 mmol)and triethylamine (11 mL, 78 mmol). The reaction mixture was stirred at0° C. for 1 hour before water (100 mL) was added. The organic layer wasseparated and extracted with 2N HCl (1×100 mL) and Brine (1×100 mL). Theorganic phase was dried over Na₂SO₄, evaporated under reduced pressure,and dried in vacuo, affording ethyl phenylmethanesulfonate (10.47 g,99%).

To a −78° C. solution of ethyl phenylmethanesulfonate (4.70 g, 23.5mmol) in anhydrous THF (100 mL) was added butyl lithium (2.5 M inhexanes, 10 mL, 25 mmol). The solution was stirred at −78° C. for 1 hourbefore crotlyl bromide (2.4 mL, 23.5 mmol) was added via syringe pumpover a 0.5-h period. The reaction mixture was stirred at −78° C. for 5hours. The mixture was warmed up to 0° C. before water (60 mL) wasslowly added. The organic layer was separated; and the aqueous layer wasextracted with EtOAc (3×60 mL). The organic layer and the extracts werecombined, dried over Na₂SO₄, and evaporated under reduced pressure,affording ethyl 1-phenylbut-3-ene-1-sulfonate (4.06 g, 70%).

A solution of ethyl 1-phenylbut-3-ene-1-sulfonate (4.06 g, 11.2 mmol) in2% TFA/CH₂Cl₂ (30 mL) was stirred at reflux for 30 hours. The solventwas evaporated under reduced pressure. The residual oil was dissolved in20% TFA/CH₂Cl₂ (30 mL). The solution was stirred at reflux for 36 hours.After cooling to room temperature, water (30 mL) was added. The organicphase was separated and extracted with a saturated solution of NaHCO₃(1×30 mL). The organic phase was dried over Na₂SO₄, evaporated todryness, and further dried in vacuo. The residual material was purifiedby flash chromatography (100% Hexanes to 80% Hexanes/Ethyl acetate),affording a mixture of sultone derivatives (1.36 g)

To ammonium hydroxide (28-30% NH₃, 30 mL, 260 mmol) was added a solutionof sultone (1.38 g, 6.1 mmol) in 1,4-dioxane (20 mL). The solution wasstirred at room temperature for 60 hours. The solvent was co-evaporatedwith EtOH. The residue was dissolved in water (30 mL), and the aqueoussolution was extracted with ethyl acetate (30 mL). The aqueous phase wasseparated, and evaporated to dryness. The solid residue was purified bypreparative HPLC; and the corresponding fractions were combined andlyophilized, affording the title compound (72 mg). ¹H NMR (D₂O, 500 MHz)δ ppm 7.29 (m, 5H), 4.07 (m, 1H), 2.99 (m, 1H), 2.35 (m, 2H), 1.49 (m,1H), 0.75 (t, 3H, J=7.3 Hz); ¹³C (D₂O, 125 MHz) δ ppm 134.80, 129.41,129.03, 128.84, 62.90, 51.68, 33.37, 25.91, 8.66; ES-MS 242 (M−1).

Preparation of 4-amino-1-phenyl-1-pentanesulfonic acid, trifluoroaceticacid salt: (Compound N31)

To a 0° C. solution of α-toluenesulfonyl chloride (10 g, 52 mmol) inanhydrous dichloromethane (200 mL) was added ethanol (6 mL, 104 mmol)and triethylamine (11 mL, 78 mmol). The reaction mixture was stirred at0° C. for 1 hour before water (100 mL) was added. The organic layer wasseparated, and extracted with 2N HCl (1×100 mL) and Brine (1×100 mL).The organic phase and extracts were combined, dried over Na₂SO₄,evaporated under reduced pressure, and dried in vacuo, affording ethylphenylmethanesulfonate (10.47 g, 99%).

To a −78° C. solution of ethyl phenylmethanesulfonate (4.70 g, 23.5mmol) in anhydrous THF (100 mL) was added butyl lithium (2.5 M inhexanes, 10 mL, 25 mmol). The mixture was stirred at −78° C. for 1 hbefore crotyl bromide (2.4 mL, 23.5 mmol) was added via syringe pumpover a 0.5 hour period. The reaction mixture was stirred at −78° C. for5 hours, and then was warmed up to 0° C. before water (60 mL) was slowlyadded. The organic layer was separated; and the aqueous layer wasextracted with EtOAc (3×60 mL). The organic layer and extracts werecombined, dried over Na₂SO₄, and evaporated under reduced pressure,affording ethyl 1-phenylbut-3-ene-1-sulfonate (4.06 g, 70%).

A solution of ethyl 1-phenylbut-3-ene-1-sulfonate (4.06 g, 11.2 mmol) in2% TFA/CH₂Cl₂ (30 mL) was stirred at reflux for 30 hours. The solventwas evaporated under reduced pressure. The residual oil was dissolved in20% TFA/CH₂Cl₂ (30 mL). The solution was stirred at reflux for 36 hours,and then cooled to room temperature, followed by addition of water (30mL). The organic phase was separated and extracted with a saturatedsolution of NaHCO₃(1×30 mL). The organic phase was dried over Na₂SO₄,evaporated to dryness, and further dried in vacuo. The residual materialwas purified by flash chromatography. (100% hexanes to 80% hexanes/Ethylacetate), affording a mixture of sultone derivatives (1.36 g)

To ammonium hydroxide (28-30% NH₃, 30 mL, 260 mmol) was added a solutionof sultone (1.38 g, 6.1 mmol) in 1,4-dioxane (20 mL). The solution wasstirred at room temperature for 60 hours. The solvent was co-evaporatedwith EtOH. The residue was dissolved in water (30 mL). The solution wasextracted with ethyl acetate (30 mL). The organic phase was recovered,dried over NaSO₄, filtered, evaporated and dried in vacuo, affording thebutanesultone (0.777 g).

To ammonium hydroxide (28-30% NH₃, 15 mL, 130 mmol) was added a solutionof the sultone derivatives obtained from step 4 (0.777 g, 3.4 mmol) in1,4-dioxane (20 mL). The solution was stirred at 85° C. for 48 hours.The solvent was co-evaporated with EtOH; and the residual solid waspurified by preparative HPLC. The corresponding fractions were combinedand lyophilized, affording the title compound (213 mg) as a pair ofenantiomers of the title compound; ¹H NMR (D₂O, 500 MHz) δ ppm 7.26 (m,5H), 3.87 (dd, 1H, J=3.6 Hz, 11.4 Hz), 3.66 (m, 1H), 2.07 (m, 2H), 1.25(m, 1H), 1.11 (m, 1H), 0.94 (d, 3H, J=6.3 Hz); ¹³C (D₂O, 125 MHz) δ ppm135.61, 129.52, 128.77, 128.34, 67.31, 66.37, 35.47, 26.32, 21.97; ES-MS243 (M−1).

Preparation of 3-(aminomethyl)bicyclo[2.2.2]oct-5-ene-2-sulfonic acid(Compound N32)

A mixture of 1,3-prop-1-ene sultone (obtained in step 3 for thepreparation of Compound N20, 720 mg, 6 mmol) and 2,3-cyclohexadiene (4.0mL, 42 mmol) in toluene (20 mL) was placed in a sealed tube and heatedat 150° C. for 30 hours. The solvent was removed and the residue wasapplied on silica gel column using hexanes/EtOAc 80/20 to 70/30 aseluant to afford the corresponding Diels-Alder adduct (1.1 g, 92%); ¹HNMR (500 MHz, CDCl₃) δ 1.30-1.62 (m, 4H), 2.76 (m, 1H), 3.00 (m, 1H),3.18 (, 1H), 3.48 (dd, J=1.0.0 & 2.0 Hz, 1H), 3.94 (dd, J=10.0 & 3.5 Hz,1H), 4.30 (t, J=8.5 Hz, 1H), 6.30 (t, J=8.5 Hz, 1H), 6.41 (t, J=8.5 Hz,1H).

A solution of the Diels-Alder adduct obtained from step 1 (1.1 g, 5.49mmol) in a solvent mixture of THF and EtOH (20 mL, v/v=1:1) was addedslowly to an aqueous solution of ammonium hydroxide (28%, 32 mL, 253mmol). The reaction mixture was stirred at room temperature for 7 hoursand concentrated under reduced pressure. The resultant solid wassuspended in EtOH, heated at reflux for 15 minutes, and cooled to roomtemperature. The solid was collected by filtration, washed with ether,and then dried, affording the title compound (700 mg, 59%); ¹H NMR (500MHz, D₂O) δ 1.10-1.28 (m, 2H), 1.40-1.58 (m, 2H), 2.40 (m, 1H), 2.56 (m,1H), 2.87-2.95 (m, 2H), 3.30 (m, 1H), 6.12 (t, J=8.0 Hz, 1H), 6.30 (t,J=8.0 Hz, 1H); ¹³C NMR (125 MHz, D₂O) δ 22.82, 25.70, 32.43, 34.28,40.72, 40.82, 62.64, 130.72, 134.49; ES-MS 216 (M−1).

Preparation of 3-(aminomethyl)bicyclo[2.2.2]octane-2-sulfonic acid(Compound N33)

Pd/C (100 mg) was added to a solution of the Diels-Alder adduct obtainedin step 1 for the preparation of3-(aminomethyl)bicyclo[2.2.2]oct-5-ene-2-sulfonic acid (Compound N32)(300 mg, 1.5 mmol) in a cosolvent of EtOAc and methanol (15 mL,v/v=2:1). The suspension was stirred under atmosphere pressure of H₂ for6 h and filtered. The filtrate was concentrated to give 300 mg of thereduced product. ¹H NMR (500 MHz, CDCl₃) δ 1.40-1.85 (m, 8H), 2.10-2.20(m, 2H), 2.88-2.92 (m, 1H), 3.38 (m, 1H), 4.28 (dd, J=10.0 & 2.0 Hz,1H), 4.41 (dd, J=10.0 & 8.0 Hz, 1H).

A solution of the reduced product prepared as described in step 1 (607mg, 3 mmol) in a solvent mixture of THF and EtOH (10 mL, v/v=1/1) wasadded slowly to an aqueous solution of ammonium hydroxide (28%, 32 mL,253 mmol). The reaction mixture was stirred at room temperature for 6hours and then concentrated. The residual solid was suspended in EtOH;the mixture was heated at reflux for 15 minutes and then cooled to roomtemperature. The solid was collected by filtration, washed with ether,and then dried, yielding the title compound (520 mg, 79%); ¹H NMR (500MHz, D₂O) δ 1.30-1.60 (m, 8H), 1.80-1.90 (m, 1H), 1.98 (m, 1H),2.22-2.29 (m, 1H), 3.10 (dd, J=13.0 & 7.0 Hz, 1H), 3.20 (d, J=10.5 Hz,1H), 3.64 (dd, J=13.0 & 7.0 Hz, 1H); ¹³C NMR (125 MHz, D₂O) δ 18.96,19.93, 25.06, 25.91, 26.94, 27.97, 36.12, 40.20, 59.69. ES-MS 218 (M−1).

Preparation of 3-amino-3-methyl-1-butanesulfonic acid (Compound N34)

To a cold (0° C.) solution of chloroethanesulfonyl chloride in MeOH (10mL) was added slowly NaOMe (25%, 4.3 g, 20 mmol). NaCl precipitatedduring the addition of NaOMe. The mixture was stirred at 0° C. for 1 h;and the cooling bath was removed. To the mixture was added2-nitropropane. pH indicated 1-2 value, and NaOMe was added until basicpH. The reaction mixture was stirred for 3 hours, and then filtered. Thefiltrate was concentrated, diluted with aqueous HCl (1 M), and thenextracted twice with EtOAc. The combined organic layers were dried overNa₂SO₄, and concentrated. The resultant residue was purified by columnchromatography using hexanes/EtOAc 80:20 to 50:50 as eluant to provide500 mg (23%) of the desired intermediate; ¹H NMR (500 MHz, CDCl₃) δ 1.65(s, 6H), 2.42 (m, 2H), 3.17 (m, 2H), 3.92 (s, 3H); ¹³C NMR (125 MHz,CDCl₃) δ 50.01, 56.17, 59.13, 66.11.

Ra-Ni (100 mg) was added to a stirred solution of the nitro intermediate(obtained from step 1, 400 mg, 1.89 mmol). The suspension was stirredunder atmospheric pressure of H₂ for 5 hours. TLC indicated completeconsumption of the starting material. The mixture was filtered and thefiltrate was concentrated to a residue. NMR of the residue showed thedesired product but contaminated probably with a sultame of cyclization.The crude product was dissolved in HCl (12N). The solution was heated atreflux for 2 hours, and then concentrated under reduced pressure toafford greenish foam. The crude was dissolved in MeOH and precipitatedby addition of Et₂O. The product was isolated by filtration, washed withMeOH and ether, to afford the title compound (140 mg, 42% overall fortwo steps); ¹H NMR (500 MHz, D₂O) δ 1.24 (s, 6H), 1.92-1.98 (m, 2H),2.82-2.89 (m, 2H); ¹³C NMR (125 MHz, D₂O) δ 24.36, 34.68, 46.04, 53.81.ES-MS 166 (M−1).

Preparation of 2-(1-aminocycohexyl)-1-ethanesulfonic acid (Compound N35)

To a cold (0° C.) solution of 2-chloroethanesulfonyl chloride (2.14 mL,20 mmol) in MeOH (10 mL) was added slowly NaOMe (25%, 4.3 g, 20 mmol).NaCl precipitated during the addition of NaOMe. The mixture was stirredat 0° C. for 1 hour; and the cooling bath was removed, followed byaddition of nitrocyclohexane (2.58 g, 20 mmol). pH indicated 1-2 value,and NaOMe was added until the mixture was basic. The reaction mixturewas stirred for 3 hours then filtered. The filtrate was concentrated anddiluted with aqueous HCl (1 M), and then extracted twice with EtOAc. Thecombined organic layers were dried over NaSO₄, and concentrated todryness. The resultant residue was purified by column chromatographyusing hexanes/EtOAc 80:20 to 50:50 as eluant. The correspondingfractions were collected and evaporated to dryness, providing thecorresponding intermediate (500 mg, 23%); ¹H NMR (500 MHz, CDCl₃) δ1.35-1.70 (m, 8H), 2.35 (m, 2H), 2.38-2.45 (m, 2H), 3.08 (m, 2H), 3.90(s, 3H). ¹³C NMR (125 MHz, CDCl₃) δ 22.36, 24.75, 34.17, 44.57, 89.88.

Ra-Ni (300 mg) was added to a stirred solution of the nitro intermediate(obtained in step 1, 1 g, 3.58 mmol). The suspension was stirred underatmospheric pressure of H₂ for 3 hours. TLC indicated completeconsumption of the starting material. The reaction was filtered and thefiltrate was concentrated. NMR of the residual material showed thedesired product but contaminated probably with the sultame ofcyclization. The crude product was dissolved in aqueous HCl (6 N, 5 mL)and the solution was heated at reflux for 2 hours, concentrated underreduced pressure to afford a foamy residue. The foamy residue wasdissolved in MeOH and the desired product was precipitated by additionof Et₂O. The product was isolated by filtration then washed with MeOHand ether to afford the title compound (422 mg, 42% overall in twosteps); ¹H NMR (500 MHz, D₂O) δ 1.20-1.32 (m, 2H), 1.32-1.56 (m, 6H),1.60-1.74 (m, 2H), 2.24 (m, 2H), 2.86 (m, 2H); ¹³C NMR (125 MHz, D₂O) δ20.82, 24.21, 31.52, 33.34, 45.06, 56.20. ES-MS 206 (M−1).

Preparation of 3-(aminomethyl)-4,5-dimethyl-1-cycloheanesulfonic acid(Compound N36)

Pd/C (100 mg) was added to a solution of6-(aminomethyl)-3,4-dimethylcyclohex-3-ene-1-sulfonic acid (420 mg, 1.92mmol) in EtOH (60% in water, 50 mL). The suspension was stirred under H₂pressure for 6 hours, filtered and concentrated to obtain 380 mg of thereduced product as a 1:1 mixture of cis/trans diastereoisomers. ¹H NMR(500 MHz, H₂O) δ 0.70 (d, J=5.5 Hz, 1.5H), 0.73 (d, J=5.5 Hz, 1.5H),0.76 (d, J=5.5 Hz, 1.5H), 0.79 (d, J=5.5 Hz, 1.5H), 1.03 (m, 1H), 1.23(m, 1H), 1.40 (m, 0.5H), 1.60 (1.78 (m, 3H), 1.85 (m, 0.5H), 2.32-2.43(m, 1H), 2.91-3.02 (m, 2H), 3.30-3.38 (m, 1H). ¹³C NMR (125 MHz, D₂O) δ14.12, 18.51, 18.90, 19.19, 25.50, 30.57, 31.18, 31.98, 33.56, 33.77,34.31, 36.87, 37.13, 38.10, 38.16, 41.01, 60.75, 60.91. ES-MS 220 (M−1).

Preparation of 3-{[(2S)-2-hydroxy-2-phenylethyl]amino}-2-propanesulfonicacid: (Compound N37)

To a solution of (S)-(+)-2-amino-1-phenylethanol (5.0 g, 36.4 mmol) in25% toluene/acetonitrile (35 mL) was added 1,3-propane sultone (4.2 g,34.7 mmol). The solution was stirred at reflux for 3 hours. The reactionmixture was cooled to room temperature. The solid was collected byfiltration and washed with acetone (2×20 mL). The solid was suspended inEtOH (40 mL). The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature, the solid material was collectedby filtration, washed with acetone (2×10 mL), and dried in a vacuum oven(50° C.), affording the title compound (7.77 g, 86%); ¹H NMR (D₂O, 300MHz) δ ppm 7.31 (m, 5H), 4.91 (dd, 1H, J=3.9 Hz, 9.3 Hz), 3.18 (m, 2H),3.12 (t, 2H, J=6.8 Hz), 2.87 (t, 2H, J=7.3 Hz), 2.02 (m, 2H); ¹³C NMR(D₂O, 75 MHz) δ ppm 139.58, 129.16, 128.97, 126.31, 69.05, 53.21, 48.07,46.60, 21.29; ES-MS 258 (M−1).

Preparation of 3-{[(2R)-2-hydroxy-2-phenylethyl]amino}-2-propanesulfonicacid: (Compound N39)

To a solution of (R)-(−)-2-amino-1-phenylethanol (5.0 g, 36.4 mmol) in25% toluene/acetonitrile (40 mL) was added 1,3-propane sultone (4.2 g,34.7 mmol). The solution was stirred at reflux for 4 hours. The reactionmixture was cooled to room temperature. The solid was collected byfiltration and washed with acetone (2×20 mL). The solid was suspended inEtOH (50 mL). The suspension was stirred at reflux for 1 hour. Themixture was cooled to room temperature, the solid material was collectedby filtration, washed with acetone (2×10 mL), and dried in a vacuum oven(50° C.), affording the title compound (8.10 g, 90%); ¹H NMR (D₂O, 300MHz) δ ppm 7.46 (m, 5H), 5.06 (dd, 1H, J=4.1 Hz, 9.0 Hz), 3.36 (m, 2H),3.29 (m, 2H), 3.02 (t, 2H, J=7.3 Hz), 2.18 (m, 2H); ¹³C NMR (D₂O, 75MHz) ppm 139.56, 129.16, 128.97, 126.12, 69.03, 53.20, 48.06, 46.59,21.27; [α]_(D)=−36.9° (c=0.0116 in water); ES-MS 260 (M+1).

Preparation of 3-[(thiophen-2-methyl)amino]-1-propanesulfonic acid(Compound N43)

To a 0° C. solution of 2-thiophene methylamine (1.0 g, 8.8 mmol) in 40%toluene/acetonitrile (15 mL) was added via syringe pump1,3-propanesultone (1.0 g, 8.4 mmol) in 40% toluene/acetonitrile (3 mL)over a 3 hour period. When the slow addition was completed, the reactionmixture was allowed to warm up to room temperature and the productstarted to precipitate. The mixture was stirred under these conditionsfor 60 hours. The solid material was collected by filtration and washedwith acetone (2×10 mL). The solid was suspended in EtOH (15 mL). Thesuspension was stirred at reflux for 1 hour. The mixture was cooled toroom temperature; and the solid material was collected by filtration,washed with acetone (2×10 mL), and dried in a vacuum oven (50° C.),affording the title compound (1.24 g, 63%). ¹H NMR (D₂O, 500 MHz) δ ppm1.99 (m, 2H), 2.85 (t, 2H, J=7.3 Hz), 3.10 (t, 2H, J=7.3 Hz), 4.35 (s,2H), 6.99 (t, 1H, J=4.4 Hz), 7.14 (d, 1H, J=2.4 Hz); ¹³C NMR (D₂O, 125MHz) δ ppm 21.39, 45.03, 45.47, 48.01, 128.00, 128.85, 131.07, 131.55;ES-MS 234 (M−1).

Preparation of 3-[1-(2-thienyl)-cyclohexylamino]-1-propanesulfonic acid(Compound N45)

To a vigorously stirred suspension of magnesium (dried, 5 g, 205 m mmol)in THF (anhydrous, 60 mL) was added slowly 2-bromothiophene (5.7 g, 35mmol) (note: exothermic reaction). The mixture was stirred for 2 hoursat room temperature, followed by dropwise addition of cyclohexanone (3g, 31 mmol) in THF (10 mL). The reaction mixture was stirred for 1 hour,quenched carefully by adding HCl (2.5N, 10 mL) and diluted with Et₂O(100 mL). The liquid part was transferred to a separator funnel to whichwas added HCl (1 M, 50 mL). The organic layer was isolated, dried(MgSO₄), and concentrated. The residual material was used in the nextstep without purification.

The residual material from step 1 was diluted in CH₂Cl₂ (60 mL). To theobtained solution was added NaN₃ (4.7 g, 73 mmol) followed by TFA (6mL). The reaction mixture was stirred for 1 hour, quenched with water,and diluted with ether. The organic layer was washed with water and 1NNH₄OH in sequence, dried over Na₂SO₄, and carefully concentrated. Theresultant azide derivative was used in the next step withoutpurification.

LAH (1 g, 26 mmol) was added portion-wise to a solution of the azidederivative (obtained from step 2) in ether (80 mL). The reaction mixturewas stirred for 3 hours before being quenched with NaOH (1N). Thequenched mixture was extracted twice with Et₂O. The combined organiclayers were dried (Na₂SO₄), and concentrated to a residue, which waspurified by column chromatography using CH₂Cl₂/MeOH 95:05 to 90:10 aseluant, affording the desired amine intermediate (2 g).

To a stirred solution of the amine intermediate (obtained in step 3, 1.5g, 8.27 mmol) in CH₃CN (25 mL) was added 1,3-propane sultone (1.01 g,8.27 mmol). The reaction mixture was stirred at room temperature overthe week-end. The solid was collected by filtration, suspended in EtOH(40 mL). The suspension was stirred at reflux for 1 hour, cooled to roomtemperature; and the solid material was collected by filtration, washedwith ethanol, and dried under high vacuum to afford the title compound(1.3 g, 78%); ¹H NMR (500 MHz, D₂O+a drop of NaOD) δ 1.10-1.28 (m, 4H),1.40 (m, 2H), 2.52 (m, 4H), 1.95 (m, 2H), 2.13 (t, J=8.0 Hz, 2H), 2.57(t, J=8.0 Hz, 2H), 6.82 (m, 2H), 7.16 (m, 1H). ¹³C NMR (125 MHz, D₂O+adrop of NaOD) δ 22.20, 24.77, 25.40, 37.10, 40.32, 49.27, 57.44, 124.46,124.63, 125.39, 126.86. ES-MS 302 (M−1).

Preparation of 3-[(2-furylmethyl)amino]-1-propanesulfonic acid (CompoundN48)

A solution of 1,3-propane sultone (1.5 g, 12.28 mmol) in CH₃CN (10 mL)was added slowly within 4 hours via a syringe pump to a boiling solutionof (2-furylmethyl)amine (6 g; 61.78 mmol) in CH₃CN (120 mL). Thereaction was stirred for an hour before being concentrated under reducedpressure. The residue was diluted with water and EtOAc. The organiclayer was discarded and the aqueous phase was washed twice with EtOAc,concentrated under high vacuum to afford the title compound as a whitesolid (2.6 g, 96%); ¹H NMR (500 MHz, D₂O) δ 1.94 (m, 2H), 2.83 (m, 2H),3.03 (m, 2H), 4.12 (s, 2H), 6.35 (m, 1H), 6.46 (m, 1H), 7.44 (m, 1H).¹³C NMR (125 MHz, D₂O) δ 21.60, 43.30, 45.54, 48.08, 11.11, 112.43,144.79, 154.27. ES-MS 218 (M−1).

Preparation of 3-[(tetrahydrofuran-2-ylmethyl)amino]-1-propanesulfonicacid (Compound N47)

A solution of 1,3-propane sultone (1.5 g, 12.58 mmol) in CH₃CN (10 mL)was added slowly within 4 h via a syringe pump to a boiling solution oftetrahydrofurfurylamine (6 g, 59.91 mmol) in CH₃CN (120 mL). Thereaction was stirred for an hour before being concentrated under reducedpressure. The residual material was diluted with water and EtOAc. Theorganic layer was discarded; and the aqueous phase was washed twice withEtOAc, concentrated under high vacuum to afford the desired product as acomplex with tetrahydrofurfurylamine. ¹H NMR (500 MHz, D₂O) δ 1.40-1.52(m, 1H), 1.75-2.00 (m, 5H), 2.71-2.88 (m, 5H), 3.62-3.75 (m, 2H), 4.00(m, 1H). ¹³C NMR (125 MHz, D₂O) δ 22.61, 25.10, 25.18, 28.38, 28.89,43.43, 47.03, 48.59, 51.69, 28.25, 28.31, 76.11, 76.89.

To the solution of product obtained in step 1 in water (20 mL) was addedone equivalent of NaOH, followed by CH₂Cl₂ (30 mL) and the mixture wasstirred vigorously. The organic layer was discarded and the aqueousphase was washed twice with CH₂Cl₂, concentrated under reduced pressureto afford the title compound (1.8 g, 96%); ¹H NMR (500 MHz, D₂O) δ 1.42(m, 1H), 1.78 (m, 4H), 1.90 (m, 1H), 2.55 (m, 4H), 2.80 (m, 2H), 3.63(m, 1H), 3.70 (m, 1H), 3.90 (m, 1H); ¹³C NMR (125 MHz, D₂O) δ 24.05,25.19, 29.12, 47.52, 49.14, 52.49, 67.96, 78.25. ES-MS 246 (M+1).

Preparation of 3-[2-(furan-2-yl)-2-propylamino]-1-propanesulfonic acid(Compound N46)

CeCl₃-7H₂O (8.00 g, 21.48 mmol) was dried at 140° C.-150° C. for 15 h.The dry solid was placed in THF (100 mL). The mixture was stirredvigorously for 1 hour, cooled to −78° C., and followed by addition ofMeLi (13.42 mL, 21.48 mmol). The suspension was stirred for 2 hours,followed by the dropwise addition of 2-furanecarbonitrile (1 g, 10.74mmol). The reaction mixture was stirred at −78° C. for 6 hours, followedby addition of concentrated aqueous NH₄OH (70 mL). The mixture waswarmed to room temperature and filtered through celite. The filtrate wasextracted with EtOAc. The organic layer was dried (Na₂SO₄) andconcentrated. The residual material was subjected to columnchromatography (CH₂Cl₂/MeOH 95:5 as the eluant) to give the desiredamine intermediate (500 mg, 36%); ¹H NMR (500 MHz, CDCl₃) δ 1.45 (s,6H), 5.60 (bs, NH₂), 6.05 (m, 1H), 6.26 (m, 1H), 7.31 (m, 1H). ¹³C NMR(125 MHz, CDCl₃) δ 29.74, 49.79, 102.28, 110.08, 141.18, 163.17.

To a stirred solution of the amine intermediate (obtained in step 1, 200mg, 1.55 mmol) in CH₃CN (10 mL) was added 1,3-propane sultone (2.16 g,17.7 mmol). The reaction mixture was stirred overnight at reflux thencooled to room temperature. The solid was collected by filtration,suspended in EtOH (40 mL). The ethanol suspension was stirred at refluxfor 1 hour, and then cooled to room temperature. The solid was collectedby filtration, washed with ethanol, and dried under high vacuum,providing the title compound (120 mg, 31%); ¹H NMR (500 MHz, D₂O) δ 1.59(m, 6H), 1.87 (m, 2H), 2.75-2.83 (m, 4H), 6.36 (m, 1H), 6.46 (m, 1H),7.45 (m, 1H). ¹³C NMR (125 MHz, D₂O) δ 21.72, 22.80, 41.18, 48.08,57.95, 110.17, 110.91, 144.36, 151.12. ES-MS 246 (M−1).

Preparation of 3-(5-indanylamino)-1-propanesulfonic acid (Compound N49)

A solution 1,3-propane sultone (8.67 g, 71 mmol) in toluene (30 mL) wasadded to a solution of 5-aminoindan (10 g, 71 mmol) in MeCN (70 mL). Themixture was heated under reflux. After 20 minutes, the mixture turnedinto a lump. Ethanol (50 mL) was added to restore stirring and thesuspension was heated at reflux for another 2 hours. The mixture wascooled to 5° C. with an ice bath. The solid was collected by suctionfiltration and rinsed with ethanol (2×20 mL). The wet cake was aspiratordried for 10 minutes. The solid was dried overnight in a vacuum oven at60° C. The resulting solid (13.57 g) was recrystallized in a mixture ofethanol (80 mL) and water (20 mL). The suspension was cooled to 1.0° C.The solid was collected by suction filtration, rinsed with ethanol (2×20mL), aspirator dried for 15 min., and further dried overnight in avacuum oven at 60° C., finishing as the title compound as a fine whitepowder (12.23 g, 67%); ¹H NMR (500 MHz, D₂O) δ 1.94 (m, 2H), 2.01 (m,2H), 2.79 (m, 4H), 2.85 (m, 2H), 3.41 (m, 2H), 7.07 (m, 1H), 7.19 (m,1H), 7.27 (m, 1H); ¹³C (125 MHz, D₂O) δ 21.07, 25.45, 32.13, 32.51,47.91, 50.53, 118.41, 120.13, 125.89, 132.66, 146.81, 147.47; ES-MS 254(M−1)

Preparation of 3-(4-indanylamino)-1-propanesulfonic acid (Compound N50)

To a 0° C. solution of 4-aminoindan (5.0 g, 37.5 mmol) in 25%toluene/acetonitrile (50 mL) was added 1,3-propane sultone (4.37 g, 35.8mmol). The mixture was stirred at reflux overnight, and then cooled toroom temperature. The solid material was collected by filtration, andwashed with acetone (2×25 mL). The resultant solid was suspended in EtOH(60 mL). The suspension was stirred at reflux for 1 hour, and thencooled to room temperature. The solid material was collected byfiltration, washed with acetone (2×25 mL) and dried in a vacuum oven(50° C.), affording the title compound (7.75 g, 85%). ¹H NMR (D₂O, 500MHz) δ ppm 2.01 (m, 4H), 2.84 (m, 6H), 3.42 (t, 2H, J=7.8 Hz), 7.05 (d1H, J=7.8 Hz), 7.18 (t, 1H, J=7.6 Hz), 7.26 (d, 1H, J=7.3 Hz); ¹³C NMR(D₂O, 125 MHz) δ ppm 21.16, 25.25, 29.87, 32.60, 47.94, 49.10, 119.84,125.81, 128.39, 137.50, 148.73; ES-MS 254 (M−1).

Preparation of3-{[2-(2-benzothiophenyl)-2-propyl]amino}-1-propanesulfonic acid(Compound N52)

A solution of benzothiophene-2-carboxaldehyde (1.5 g, 9.2 mmol),hydroxylamine hydrochloride (760 mg, 11.0 mmol) inN-methyl-2-pyrrolidinone (NMP, 15 mL) was stirred at 115° C. for 4hours. After the reaction mixture was cooled to room temperature, and itwas poured into water (50 mL). The resultant mixture was extracted withdiethyl ether (2×20 mL). The organic extracts were combined, dried overNa₂SO₄, evaporated to dryness, and further dried in vacuo. The residualmaterial was purified by flash chromatography (R_(f)=0.45, 10%EtOAc/hexanes), affording 1-benzothiophene-2-carbonitrile (0.850 g,58%).

Cerium chloride heptahydrate (7.5 g, 20.2 mmol) was dried in vacuo at150° C. overnight, and then placed in anhydrous THF (40 mL). Thesuspension was stirred at room temperature and sonicated for 15 minutes.The mixture was cooled to −50° C. before methyl lithium (1.6 M in Et2O,12.6 mL, 20.2 mmol) was slowly added. The mixture was stirred at −50° C.for 1 h, followed by addition of 1-benzothiophene-2-carbonitrile (fromstep 1, 0.850 g, 5.3 mmol) via syringe. The reaction mixture was stirredat −50° C. for 2 hours, and then quenched with concentrated NH₄OH (15mL). The mixture was warmed up to 0° C. before the solid material wasremoved by filtration. The organic phase was separated and the solventwas evaporated. The residue was dissolved in diethyl ether (25 mL). Thesolution was extracted with Brine (2×25 mL). The organic phase was driedover Na₂SO₄. After removal of the solvent by evaporation, the residuewas dried in vacuo, purified by flash chromatography (R_(f)=0.33, 5%MeOH/CH₂Cl₂, affording 2-(2-benzothiophenyl)-2-propylamine (0.620 g,61%).

To a solution of 2-(2-benzothienyl)-2-propylamine (0.620 g, 3.2 mmol) in25% toluene/acetonitrile (15 mL) was added 1,3-propane sultone (0.377 g,3.1 mmol). The reaction mixture was stirred at reflux overnight. Thesolid material was collected by filtration and washed with acetone (2×10mL). The solid was recrystallized in EtOH (10 mL) and water (2 mL). Theresultant light-blue crystals were dissolved in a hot solvent mixture(15% water/EtOH); and the solution was treated with activated charcoal.The hot suspension was filtered on celite. The filtrate was evaporatedto dryness. The resulting solid was suspended in acetone (20 mL),collected by filtration and dried in a vacuum oven (50° C.), affordingthe title compound (0.514 g, 51%). ¹H NMR (DMSO, 500 MHz) δ ppm 1.77 (s,6H), 1.92 (m, 2H), 2.59 (t, 2H, J=6.3 Hz), 2.90 (m, 2H), 7.40 (m, 2H),7.59 (s, 1H), 7.85 (m, 1H), 7.95 (m, 1H), 9.38 (s (br), 2H); ¹³C NMR(DMSO, 125 MHz) δ ppm 144.66, 139.60, 139.55, 125.97, 125.58, 124.89,123.26, 59.04, 50.07, 42.73, 26.62, 22.78; ES-MS 312 (M−1).

Preparation of 3-(4-chlorophenyl-2-propylamino)-1-propanesulfonic acid(Compound N54)

CeCl₃-7H₂O (20.15 g, 59.1 mmol) was dried at 150° C. for 15 hours. Thedry solid was placed in THF (200 mL). The mixture was stirred vigorouslyfor 1.5 hours, and cooled to −78° C. To the suspension was added MeLi(1.6 M, 37 mL, 59.2 mmol). The suspension was warmed to −50° C., stirredfor 1 hour, and then cooled to −78° C., followed by the dropwiseaddition of a solution of 4-chlorothiobenzamide (2.0 g, 11.6 mmol) inTHF (15 mL). The mixture was warmed slowly to 0° C. in 2.5 hours, andthen was cooled to −50° C., followed by addition of concentrated aqueousNH₄OH (50 mL). The mixture was warmed to room temperature and filteredthrough celite. The filtrate was extracted with EtOAc; the organic layerwas dried (Na₂SO₄) and concentrated to dryness. The desired aminederivative (0.80 g, 41%) was obtained after purification using columnchromatography on silica gel (CH₂Cl₂/MeOH as the eluant).

To a stirred solution of the amine derivative (obtained in step 1, 0.8g, 4.7 mmol) in a solvent mixture of CH₃CN (10 mL) and toluene (3 mL)was added 1,3-propane sultone (600 mg, 5 mmol). The reaction mixture wasstirred overnight at reflux and then cooled to room temperature. Thesolid material was collected by filtration, suspended in EtOH (10 mL),and stirred at reflux for 1 hour. The suspension was then cooled to roomtemperature; and the solid was collected by filtration, washed withethanol, and dried under high vacuum to afford the title compound (1.11g, 81%); ¹H NMR (500 MHz, DMSO-d6) δ 1.66 (s, 6H), 1.91-1.95 (m, 2H),2.61 (t, J=6.3 Hz, 2H), 2.75 (br s, 2H), 7.54 (d, J=8.8 Hz, 2H), 7.60(d, J=8.8 Hz, 2H), 9.21 (br s, 2H); ¹³C (125 MHz, DMSO-d6) δ 22.13,25.09, 41.86, 49.31, 59.73, 128.23, 128.82, 133.37, 138.56; ES-MS 290(M−1)

Preparation of 3-[(1-thien-2-ylethyl)amino]-1-propanesulfonic acid(Compound N55)

To a solution of 1-thiophene-2-yl ethylamine (0.500 g, 3.9 mmol) in 25%toluene/acetonitrile (15 mL) was added 1,3-propane sultone (0.457 g, 3.7mmol). The reaction mixture was stirred at reflux for 3 hours. Aftercooling to room temperature, the solid material was collected byfiltration and washed with acetone (2×10 mL). The solid product wassuspended in EtOH (20 mL). The mixture was stirred at reflux for 1 hour.After cooling to room temperature, the solid material was collected byfiltration, washed with acetone (2×10 mL) and dried in a vacuum oven(50° C.), affording the title compound (0.556 g, 60%). ¹H NMR (DMSO, 500MHz) δ ppm 1.57 (d, 3H, J=6.8 Hz), 1.91 (m, 2H), 2.59 (t, 2H, J=6.8 Hz),2.86 (m, 1H), 3.15 (m, 1H), 4.71 (m, 1H), 7.09 (t, 1H, J=3.9 Hz), 7.26(d, 1H, J=2.9 Hz), 7.62 (d, 1H, J=4.9 Hz), 9.11 (s (br), 2H); ¹³C NMR(DMSO, 125 MHz) δ ppm 20.24, 22.43, 45.39, 49.91, 52.32, 128.00, 128.94,139.80; ES-MS 248 (M−1).

Preparation of 3-(4-fluorophenyl-2-propylamino)-1-propanesulfonic acid(Compound N57)

CeCl₃-7H₂O (21.5 g, 57.7 mmol) was dried at 150° C. for 15 hours. To thesolid was added THF (250 mL). The mixture was stirred vigorously for 1.5hours, cooled to −78° C. To the suspension was added MeLi (1.6 M, 38 mL,60.8 mmol). The suspension was warmed to −50° C., stirred for 1 hour andthen cooled to −78° C., followed by the dropwise addition of4-fluorothiobenzamide (2.2 g, 14 mmol). The mixture was warmed slowly to0° C. in 2.25 hours, and then was cooled to at −50° C., followed byaddition of concentrated aqueous NH₄OH (40 mL). The mixture was warmedto room temperature and filtered through celite. The filtrate wasextracted with EtOAc; and the organic layer was dried (Na₂SO₄) andconcentrated. The resultant residue was separated using columnchromatography (CH₂Cl₂/MeOH as the eluant), yielding the correspondingamine derivative (0.78 g, 36%).

To a stirred solution of the amine derivative (obtained in step 1, 780mg, 5.09 mmol) in mixed solvents of CH₃CN (7 mL) and toluene (3 mL) wasadded 1,3-propane sultone (650 mg, 5.10 mmol). The reaction mixture wasstirred overnight at reflux then cooled to room temperature. The solidwas collected by filtration, suspended in EtOH (7 mL) and stirred atreflux for 1 hour. The suspension was then cooled to room temperature;and the solid was collected by filtration, washed with ethanol and driedunder high vacuum to afford the title compound (1.16 g, 83%); ¹H NMR(500 MHz, DMSO-d6) δ 1.65 (s, 6H), 1.91 (qt, J=6.3 Hz, 2H), 2.60 (t,J=6.3 Hz, 2H), 2.75 (br s, 2H), 7.30-7.33 (m, 2H), 7.60-7.63 (m, 2H),9.21 (br s, 2H); ¹³C (125 MHz, DMSO-d6) δ 22.05, 25.22, 41.89, 49.46,59.60, 115.63 (d, J=21.1 Hz), 128.56 (d, J=8.6 Hz), 135.83, 161.91 (d,J=245 Hz); ¹⁹F (282 MHz, DMSO-d6) δ −114.0 to −114.1 (m); ES-MS 274(M−1).

Preparation of3-{[1-methyl-1-(5-methylthien-2-yl)ethyl]amino}-1-propanesulfonic acid:(Compound N60)

A solution of 5-methyl-2-thiophenecarboxaldehyde (3.0 g, 23.8 mmol),hydroxylamine hydrochloride (2.0 g, 28.7 mmol) inN-methyl-2-pyrrolidinone (NMP, 40 mL) was stirred at 115° C. for 4hours. After cooling to room temperature, the solution was poured inwater (50 mL) and extracted with diethyl ether (3×40 mL). The organicextracts were combined, dried over Na₂SO₄. Solvent was evaporated; andthe residue was dried in vacuo, and purified by flash chromatography0.41, 10% EtOAc/hexanes), affording the pure5-methylthiophene-2-carbonitrile (1.30 g, 44%).

Cerium chloride heptahydrate (10 g, 26.9 mmol) was dried in vacuo at150° C. overnight. To this solid was added anhydrous THF (100 mL). Thesuspension was stirred at room temperature for 30 minutes. The reactionmixture was cooled to −50° C. before methyl lithium (1.6 M in Et₂O, 26.9mL, 26.9 mmol) was slowly added. The mixture was stirred at −50° C. for1 hour, and then cooled to −78° C., followed by addition of5-methylthiophene-2-carbonitrile obtained from step 1 (1.3 g, 10.6 mmol)via syringe. The reaction mixture was stirred at −50° C. for 2 hours,and then quenched with concentrated NH₄OH (30 mL). The mixture waswarmed up to 0° C., and the solid material was removed by filtration.The organic phase was evaporated to dryness. The residue was dissolvedin diethyl ether (30 mL). The solution was extracted with brine (2×30mL). The combined organic phase was dried over Na₂SO₄ and the solventwas evaporated. The residue was dried in vacuo, and purified by flashchromatography (R_(f)=0.48, 5% MeOH/CH₂Cl₂), affording1-methyl-1-(5-methylthien-2-yl)ethylamine (920 mg, 61%).

To a solution of 1-methyl-1-(5-methylthien-2-yl)ethylamine (920 mg, 6.5mmol) in 25% toluene/acetonitrile (15 mL) was added 1,3-propane sultone(758 mg, 6.2 mmol). The reaction mixture was stirred at refluxovernight, and cooled to room temperature. The solid material wascollected by filtration, washed with acetone (2×15 mL), and suspended inEtOH (20 mL). The suspension was stirred at reflux for 1 hour, and thencooled to room temperature. The solid material was collected byfiltration, washed with acetone (2×10 mL) and dried in a vacuum oven(50° C.), affording the title compound (1.26 g, 74%). ¹H NMR (D₂O, 500MHz) δ ppm 1.82 (s, 6H), 1.92 (m, 2H), 2.48 (s, 3H), 2.90 (t, 2H, J=7.3Hz), 3.00 (t, 2H, J=7.8 Hz), 6.87 (m, 1H), 7.09 (d, 1H, J=3.4 Hz); ¹³CNMR (D₂O, 125 MHz) δ ppm 14.48, 21.77, 25.66, 41.03, 48.10, 59.71,125.81, 128.02, 139.53, 142.84; ES-MS 276 (M−1).

Preparation of 4-amino-1-hydroxy-1-(5-methylthien-2-yl)-2-butanesulfonicacid (Compound N63)

To a −78° C. solution of 1,3-propane sultone (2.5 g, 20.5 mmol) inanhydrous THF (100 mL) was added butyl lithium (2.5 M in hexanes, 9 mL,22.5 mmol). The solution was stirred at −78° C. for 0.5 hours, followedby addition of 5-methyl-2-thiophenecarboxaldehyde (2.2 mL, 20.5 mmol)via syringe pump over a 0.5 hour period. The reaction mixture wasstirred at −78° C. for 3 hours, and then warmed up to 0° C., followed bya slow addition of water (50 mL). The organic layer was separated; andthe aqueous layer was extracted with Et₂O (2×50 mL). The organic phaseand extracts were combined and dried over Na₂SO₄. Solvent was removedunder reduced pressure. The residue was purified on a flashchromatography (Rf=0.26, 70% Hex/EtOAc), affording the correspondingsultone derivative (2.08 g, 41%).

To 0° C. concentrated ammonium hydroxide (28-30% NH₃, 50 mL) was added asolution of sultone (3.04 g, 13.3 mmol) in THF (15 mL) via syringe pumpover a 4 hour period. The solution was stirred at 0° C. for 1 h and atroom temperature overnight. The solvent was co-evaporated with EtOH. Thesolid was suspended in EtOH (15 mL). The mixture was stirred at refluxfor 1 hour. After cooling to room temperature, the solid was filtered,washed with acetone (2×10 mL), and dried in a vacuum oven (50° C.),affording the title compound as a mixture of diastereoisomers (0.785 g,35%). ¹H NMR (D₂O, 500 MHz) ppm 6.80 (d, 0.2H, J=3.4 Hz), 6.72 (d, 0.8H,J=2.9 Hz), 6.65 (m, 1H), 5.43 (d, 0.8H, J=2.4 Hz), 5.12 (d, 0.2H, J=7.3Hz), 3.08 (m, 1.6H), 2.97 (m, 0.4H), 2.85 (m, 1H), 2.31 (m, 3H), 2.04(m, 1.6H), 1.84 (m, 0.4 H); ¹³C (D₂O, 125 MHz) δ ppm 14.44, 22.85,38.54, 63.80, 68.70, 124.18, 125.36, 140.26, 142.98; ES-MS 264 (M−1).

Preparation of3-(4-trifluoromethylphenyl-2-propylamino)-1-propanesulfonic acid(Compound N64)

CeCl₃-7H₂O (21.0 g, 56.4 mmol) was dried at 150° C. for 15 hours. To thesolid was added THF (250 mL). The mixture was stirred vigorously for 1.5hours, cooled to −78° C. To the suspension was added MeLi (1.6 M, 38 mL,60.8 mmol). The suspension was warmed to −50° C., stirred for 1 hour andthen cooled to −78° C., followed by dropwise-addition of a solution of4-trifluoromethylthiobenzamide (2.46 g, 12 mmol) in THF (20 mL). Themixture was warmed slowly to 0° C. in 2.5 h, and then was cooled to −50°C. followed by addition of concentrated aqueous NH₄OH (70 mL). Themixture was warmed to room temperature and filtered through celite. Thefiltrate was extracted with EtOAc; and the organic layer was dried(Na₂SO₄) and concentrated. The residue was separated using columnchromatography (CH₂Cl₂/MeOH as the eluant), affording the correspondingamine (1.69 g, 69%).

To a stirred solution of the amine (obtained in step 1, 1.69 g, 8.3mmol) in mixed solvents of CH₃CN (10 mL) and toluene (3 mL) was added1,3-propane sultone (0.75 mL, 8.5 mmol). The reaction mixture wasstirred overnight at reflux and then cooled to room temperature. Thesolid was collected by filtration and suspended in EtOH (10 mL). Thesuspension was stirred at reflux for 1 hour, and then cooled to roomtemperature. The solid material was collected by filtration, washed withethanol and dried under high vacuum to give the title compound, 2.39 g(89%); ¹H NMR (300 MHz, DMSO-d6) δ 1.69 (s, 6H), 1.94 (qt, J=6.4 Hz,2H), 2.60 (t, J=6.4 Hz, 2H), 2.79 (br s, 2H), 7.82 (q, J=8.5 Hz, 4H),9.32 (br s, 2H); ¹³C (75 MHz, DMSO-d6) δ 22.25, 25.15, 41.97, 49.26,59.93, 123.70 (q, J=271 Hz), 125.52 (d, J=3.5 Hz), 126.89, 128.71 (q,J=31.9 Hz), 143.84; ¹⁹F (282 MHz, DMSO-d6) δ −61.87 (s); ES-MS 324 (MA)

Preparation of 3-(2-phenyl-2-butylamino)-1-propanesulfonic acid(Compound N65)

The flask was closed with a septum and connected to a 20% NaOH scrubber.

Sodium cyanide (powdered, 2.6 g) was added in portions to acetic acid(10 mL). The mixture was stirred for 10 minutes at room temperature. Asolution of sulfuric acid (8 mL) in acetic acid (10 mL) was addeddropwise over a 20 minute period. Then, the 2-phenyl-2-butanol (5 g,33.3 mmol) was added dropwise over 5 min. The mixture was stirred atroom temperature for 22 hours then cooled to 0° C. with an ice-waterbath. The pH of the solution was adjusted to 9 with addition of ammoniumhydroxide (460 mL). The organic phase was separated and the aqueouslayer was extracted with ether (3×30 mL). The organic phase and extractswere combined, washed with saturated potassium carbonate (1×5 mL), anddried over sodium sulfate. The solvent was evaporated under reducedpressure. The residue was purified by flash chromatography on silica gel(MeOH/CH₄Cl₂ as eluant), affording clear yellow oil (3.84 g, 65%).

A solution of NaOH (20%, 30 mL) was added to the crude product from step1 (3.84 g). The mixture was heated at reflux for 2 h, and then cooled toroom temperature. Sodium chloride (7.5 g) was added to facilitate thephase separation. The organic layer were separated and the aqueous layerwas extracted with mixed solvent of toluene and MTBK (3×5 mL, v/v=1:3).The combined organic layers were washed with brine (1×5 mL), dried oversodium sulfate and filtered. The filtrate was used in the next stepwithout purification.

A solution of 1,3-propane sultone (400 mg, 3.3 mmol) and2-phenyl-2-aminobutane (425 mg, 2.85 mmol, from step 2) in mixedsolvents of toluene and CH₃CN (5 mL, v/v=3:7) was heated under refluxfor 22 h and then cooled to room temperature. The product had formed agum. It was triturated with ether/ethanol to provide a brownish solid.The solid was collected by suction filtration. The crude solid waspurified by reverse phase preparative HPLC (299 mg, 39%); ¹H NMR (500MHz, D₂O) δ 0.77 (t, J=7.3 Hz, 3H), 1.77 (s, 3H), 1.99-2.11 (m, 3H),2.20-2.27 9 m, 1H), 2.66-2.71 (m, 1H), 2.79-2.89 (m, 2H), 3.06-3.11 (m,1H), 7.42-7.45 (m, 1H), 7.49-7.55 (m, 4H); ¹³C (125 MHz, D₂O) δ 8.62,19.90, 23.37, 34.22, 43.15, 50.29, 66.31, 127.93, 130.38, 130.60,138.16; ES-MS 270 (M−1):

Preparation of 3-(4-methoxyphenyl-2-propylamino)-1-propanesulfonic acid(Compound N66)

CeCl₃-7H₂O (21.5 g, 57.7 mmol) was dried at 150° C. for 15 hours. To thesolid was added THF (250 mL). The mixture was stirred vigorously for 1.5hours, and then cooled to −78° C. To the suspension was added MeLi (1.6M, 38 mL, 60.8 mmol). The suspension was warmed to −50° C., stirred for1 hour and then cooled to −78° C., followed by dropwise-addition of asolution of 4-trifluoromethylthiobenzamide (2.00 g, 12 mmol) in THF (20mL). The mixture was warmed slowly to −50° C. in 4 hours. Concentratedaqueous NH₄OH (70 mL) was added and the mixture was warmed to roomtemperature and filtered through celite. The filtrate was extracted withEtOAc, the organic layer dried (Na₂SO₄) and concentrated. The residuewas separated using column chromatography (CH₂Cl₂/MeOH as the eluant),yielding the corresponding amine (0.60 g, 30%).

To a stirred solution of the amine (obtained in step 1, 0.60 mg, 3.6mmol) in mixed solvents of CH₃CN (10 mL) and toluene (2 mL) was added1,3-propane sultone (0.34 mL, 3.8 mmol). The reaction mixture wasstirred overnight at reflux then cooled to room temperature. The solidwas collected by filtration, suspended in EtOH (10 mL). The suspensionwas stirred at reflux for 1 hour, and then cooled to room temperature.The solid was collected by filtration, washed with ethanol and driedunder high vacuum to afford the title compound, 0.97 g (94%); ¹H NMR(300 MHz, DMSO-d6) δ 1.64 (s, 6H), 1.91 (qt, J=6.5 Hz, 2H), 2.58 (t,J=6.3 Hz, 2H), 2.71 (br s, 2H), 3.77 (s, 3H), 6.99-7.02 (m, 2H),7.47-7.50 (m, 2H), 9.06 (br s, 2H); ¹³C NMR (75 MHz, DMSO-d6) δ 22.09,25.27, 41.77, 49.40, 55.20, 59.67, 114.04, 127.54, 131.34, 159.21; ES-MS286 (M−1).

Preparation of 3-(3-chlorophenyl-2-propylamino)-1-propanesulfonic acid(Compound N67)

CeCl₃-7H₂O (21.5 g, 57.7 mmol) was dried at 150° C. for 15 hours. To thesolid was added THF (250 mL). The mixture was stirred vigorously for 1.5hours, cooled to −78° C., and to the suspension was added MeLi (1.6 M,40 mL, 64 mmol). The suspension was warmed to −50° C., stirred for 1hour and then cooled to −78° C., followed by dropwise-addition of asolution of 3-chlorobenzonitrile (2.5 g, 18 mmol) in THF (20 mL). Themixture was warmed slowly to 0° C. in 2.5 hours, and then was cooled toat −50° C. Concentrated aqueous NH₄OH (45 mL) was added and the mixturewas warmed to room temperature and filtered through celite. The filtratewas extracted with EtOAc, the organic layer dried (Na₂SO₄) andconcentrated. The residue was separated using column chromatography(CH₂Cl₂/MeOH as the eluant) to yield the corresponding amine (2.1 g,69%).

To a stirred solution of the amine (obtained in step 1, 2.1 g, 12.4mmol) in mixed solvents of CH₃CN (12 mL) and toluene (3 mL) was added1,3-propane sultone (1.6 g, 13 mmol). The reaction mixture was stirredovernight at reflux then cooled to room temperature. The solid wascollected by filtration, suspended in EtOH (20 mL). The suspension wasstirred at reflux for 1 hour, and then cooled to room temperature. Thesolid was collected by filtration, washed with ethanol and dried underhigh vacuum to afford the title compound (3.1 g, 86%); ¹H NMR (500 MHz,DMSO-d6) δ 1.66 (s, 6H), 1.93 (qt, J=6.5 Hz, 2H), 2.64 (t, J=6.6 Hz,2H), 2.78 (br s, 2H), 7.48-7.57 (m, 3H), 7.63 (s, 1H), 9.25 (br s, 2H);¹³C (100 MHz, DMSO-d6) δ 22.13, 25.02, 41.95, 49.34, 59.86, 124.90,126.31, 128.63, 130.79, 133.69, 142.10; ES-MS 290, 292 (M−1).

Preparation of 4-[(1R)-indan-1-ylamino]-2-butanesulfonic acid: (CompoundN69)

To a solution of (R)-(−)-1-aminoindan (2.0 g, 15.0 mmol) in 25%toluene/acetonitrile (15 mL) was added 2,4-butane sultone (1.95 g, 14.3mmol). The reaction mixture was stirred at reflux for 3 hours, and thencooled to room temperature. The solid material was collected byfiltration and washed with acetone (2×15 mL). The solid was suspended inEtOH (20 mL). The suspension was stirred at reflux for 1 hour, and thencooled to room temperature. The solid material was collected byfiltration, washed with acetone (2×10 mL) and dried in a vacuum oven(50° C.), affording the title compound (3.04 g, 79%). ¹H NMR (D₂O, 500MHz) δ ppm 1.14 (d, 1.5H, J=2.9 Hz), 1.16 (d, 1.5H, J=2.9 Hz), 1.78 (m,1H), 2.09 (m, 2H), 2.40 (m, 1H), 2.92 (m, 2H), 3.00 (m, 1H), 3.12 (t,2H, J=8.0 Hz), 4.67 (m, 1H), 7.21 (m, 1H), 7.28 (m, 2H), 7.38 (d, 1H,J=7.8 Hz); ¹³C NMR (D₂O, 125 MHz) δ ppm 14.74, 14.82, 28.29, 28.37,28.64, 28.65, 29.83, 43.03, 43.09, 53.28, 53.31, 62.91, 62.95, 125.52,125.71, 127.19, 130.28, 136.44, 145.34; [α]_(D)=−0.5° (c=0.0083 inwater); ES-MS 268 (M−1).

Preparation of 4-[(1S)-indan-1-ylamino]-2-butanesulfonic acid: (CompoundN70)

To a solution of (S)-(+)-1-aminoindan (2.0 g, 15.0 mmol) in 25%toluene/acetonitrile (15 mL) was added 2,4-butane sultone (1.95 g, 14.3mmol). The reaction mixture was stirred at reflux for 3 h, and thencooled to room temperature. The solid material was collected byfiltration, washed with acetone (2×15 mL), and then was suspended inEtOH (20 mL). The suspension was stirred at reflux for 1 hour, andcooled to room temperature. The solid material was collected byfiltration, washed with acetone (2×10 mL) and dried in a vacuum oven(50° C.), affording the title compound (3.32 g, 86%). ¹H NMR (D₂O, 500MHz) δ ppm 1.16 (m, 3H), 1.78 (m, 1H), 2.10 (m, 2H), 2.42 (m, 1H), 2.86(m, 2H), 3.01 (m, 1H), 3.13 (t, 2H, J=7.8 Hz), 4.68 (m, 1H), 7.22 (m,1H), 7.29 (m, 2H), 7.39 (d, 1H, J=7.8 Hz); ¹³C NMR (D₂O, 125 MHz) δ ppm14.74, 14.82, 28.29, 28.37, 28.64, 28.65, 29.83, 43.03, 43.09, 53.28,53.31, 62.91, 62.95, 125.51, 125.70, 127.19, 130.28, 136.43, 145.34;[α]_(D)=+0.8° (c=0.0126 in water); ES-MS 268 (M−1).

Preparation of 4-amino-1-(1-benzothien-2-yl)-2-butanesulfonic acid(Compound N71)

Sodium borohydride (250 mg, 6.5 mmol) was added in two portions to a 0°C. solution of benzo[b]thiophene-2-carboxaldehyde (2.0 g, 12.3 mmol) inethanol (15 mL). The reaction mixture was stirred at room temperaturefor 2 hours. The volume of solvent was reduced to ⅓ by evaporation.Diethyl ether (20 mL) and water (20 mL) were added. The organic layerwas separated and the aqueous phase was extracted with diethyl ether(2×20 mL). The organic layer and extracts were combined and dried overNa₂SO₄. Solvent was evaporated under reduced pressure, affording1-benzothien-2-ylmethanol (2.02 g, 99%).

To a 0° C. solution of 1-benzothien-2-ylmethanol (2.02 g, 12.3 mmol) inanhydrous CH₂Cl₂ (25 mL) was added phosphorus tribromide (1.7 mL, 18.4mmol). The reaction mixture was stirred at room temperature for 1 hour.After the mixture was cooled to 0° C., water (20 mL) was slowly added.The organic phase was separated and the aqueous phase was extracted withCH₂Cl₂ (2×20 mL). The extracts were combined with the organic phase anddried over Na₂SO₄. Solvent was evaporated under reduced pressure,affording 2-(bromomethyl)-1-benzothiophene (2.57 g, 92%).

To a −78° C. solution of 1,3-propane sultone (1.38 g, 11.3 mmol) inanhydrous THF (100 mL) was added butyl lithium (2.5 M in hexanes, 5.0mL, 12.5 mmol). The solution was stirred at −78° C. for 0.5 hours,followed by addition of 2-(bromomethyl)-1-benzothiophene (2.57 g, 11.3mmol, from step 2) via syringe pump over a 0.5-h period. The reactionmixture was stirred at −78° C. for 3 hours and then warmed up to 0° C.before water (100 mL) was slowly added. The organic layer was separatedand the aqueous layer was extracted with Et₂O (2×100 mL). The organiclayer and extracts were combined, dried over Na₂SO₄, and evaporatedunder reduced pressure. The residue was purified by a flashchromatography (Rf=0.15, 70% Hex/EtOAc), affording the correspondingsultone (970 mg, 32%).

To a 0° C. aqueous solution of ammonium hydroxide (28-30%, 50 mL) inacetone (10 mL) was added via syringe pump over a 4 hour period asolution of the sultone (3.04 g, 13.3 mmol, from step 3) in acetone (15mL). The solution was stirred at 0° C. for 1 hour and at roomtemperature for 3 hours. The solvent was co-evaporated with EtOH. Thesolid was suspended in acetone (20 mL), and then collected byfiltration, washed with acetone (1×10 mL), and dried in a vacuum oven(50° C.), affording the title compound (0.390 g, 38%); ¹H NMR (DMSO, 500MHz) δ ppm 1.81 (m, 2H), 2.81 (m, 1H), 2.90 (m, 2H), 3.00 (m, 1H), 3.48(m, 1H), 7.28 (m, 3H), 7.61 (s (broad), 2H), 7.71 (d, 1H, J=7.8 Hz),7.88 (d, 1H, J=7.8 Hz); ¹³C NMR (DMSO, 125 MHz) δ ppm 27.90, 32.81,38.38, 59.05, 122.95, 123.23, 123.60, 124.46, 125.00, 139.58, 140.54,144.08; ES-MS 284 (M−1).

Preparation of3-{[3-(methoxycarbonyl)thien-2-yl]amino}-1-propanesulfonic acid:(Compound N72)

To a solution of methyl 2-amino-3-thiophenecarboxylate (3.0 g, 19.1mmol) in 25% toluene/acetonitrile (20 mL) was added 1,3-propane sultone(2.22 g, 18.2 mmol). The reaction mixture was stirred at reflux for 5hours, and then cooled to room temperature. The reaction mixture wasconcentrated under reduced pressure. The oil residue was extracted withdiethyl ether (1×30 mL) and water (1×30 mL). The residual material fromaqueous phase was purified by preparative HPLC, affording the titlecompound (0.55 g, 10%). ¹H NMR (D₂O, 500 MHz) δ ppm 1.91 (m, 2H), 2.82(t, 2H, J=7.6 Hz), 3.36 (t, 2H, J=7.1 Hz), 3.70 (s, 3H), 6.84 (m, 1H),7.56 (d, 1H, J=5.4 Hz); ¹³C NMR (D₂O, 125 MHz) δ ppm 23.76, 45.55,48.26, 52.25, 119.36, 134.48, 149.98, 165.46; ES-MS 278 (M−1).

Preparation of 4-amino-1-dibenzyl-2-butanesulfonic acid (Compound N75)

To a −78° C. solution of 1,3-propane sultone (5.0 g, 41 mmol) inanhydrous THF (150 mL) was added butyl lithium (2.5 M in hexanes, 18 mL,45 mmol). The solution was stirred at −78° C. for 0.5 h before benzylbromide (4.9 mL, 41 mmol) was added via syringe pump over a 0.5-hperiod. The reaction mixture was stirred at −78° C. for 2 hours. Thereaction mixture was warmed up to 0° C. before water (100 mL) was slowlyadded. The organic layer was collected. The aqueous phase was extractedEtOAc (2×25 mL). The organic layer and the extracts were combined, driedover Na₂SO₄, and evaporated under reduced pressure. The residue waspurified by flash chromatography (Rf=0.25, 80% Hex/EtOAc), affording1,1-dibenzyl-1,3-propane sultone (0.637 g).

To a 0° C. solution of aqueous solution of ammonium hydroxide (28-30%,10 mL) in acetone (5 mL) was added via syringe pump over a 4 hour perioda solution of 1,1-dibenzyl-1,3-propane sultone (0.636 g, 2.1 mmol) inacetone (10 mL). The solution was stirred at room temperature overnight.The solvent was co-evaporated with EtOH. The resultant solid wassuspended in acetone (20 mL), collected by filtration, and dried in avacuum oven (50° C.), affording the title compound (0.420 g, 63%). ¹HNMR (500 MHz, DMSO) δ (ppm) 1.67 (t, 2H, J=6.8 Hz), 2.69 (d, 2H, J=14.2Hz), 3.15 (m, 4H), 7.18 (m, 10H), 7.61 (s (br), 2H); ¹³C NMR (125 MHz,DMSO) δ (ppm) 30.39, 35.61, 40.57, 60.89, 125.95, 127.46, 131.36,137.71; ES-MS 318 (M−1).

Preparation of 3-amino-2-(thien-2-ylmethyl)propane-1-sulfonic acid (twomethods) (Compound N83) Method 1:

To a cold (−78° C.) solution of 3-hydroxypropionitrile (2 g, 28.12 mmol)in THF (60 mL) was added a solution of lithium bis(trimethylsilyl)amide(1 M in THF, 56 mL). After the reaction mixture was stirred for 1 h atthis temperature, 2-thieylmethyl bromide (4.98 g, 28.12 mml) was addeddropwise. The reaction mixture was left warming to reach 0° C. at whichtemperature the mixture was stirred for 2 hours. The reaction wasquenched with 1N HCl and extracted with EtOAc. The organic layer waswashed with 1N HCl, dried over Na₂SO₄ and concentrated. The residue wasapplied on silica gel column (eluant: hexane:EtOAc 70:30 to 50:50) toafford 3-hydroxy-2-(2-thieylmethyl)-1-propionitrile (2.0 g, 42%); ¹H NMR(500 MHz, CDCl₃) δ 2.60 (bs, 1H), 3.00 (m, 1H), 3.20 (m, 2H), 3.80 (m,2H). 6.97 (m, 2H), 7.22 (m, 1H); The dialkylated product was isolated in23% yield (1.7 g).

To a stirred solution of 3-hydroxy-2-(2-thieylmethyl)-1-propionitrile(obtained in step 1, 1 g, 6 mmol) in THF (60 mL) was added portion-wiseLAH (450 mg, 12 mmol). The reaction was stirred for 2 h, quenched withNaOH (1 M) and left stirred vigorously for 1 hour before the addition ofBoc₂O (1.6 g, 7.2 mmol). The reaction mixture was stirred for 2 hoursand then diluted with Et₂O. The two phases were separated and theorganic layer was dried and concentrated. The residual material waspurified by column chromatography (Hexanes/EtOAc 70:30 as eluant) toafford the corresponding alcohol product (1.43 g, 88% yield).

To a cold (0° C.) solution of the alcohol product of step 2 (630 mg,2.32 mmol) in CH₂Cl₂ (30 mL) was added NEt₃ (646 μL, 4.64 mmol) followedby MsCl (200 μL, 2.55 mmol). The reaction mixture was stirred for 1hour, diluted with H₂O. The organic layer was isolated and concentratedto give the corresponding mesylate which was used in the next stepwithout further purification.

The solution of mesylate (obtained in step 3) in EtOH (5 mL) was addeddropwise to a refluxed solution of Na₂SO₃ (440 mg, 3.48 mmol). After 15minutes, the starting material was completely consumed. The reactionmixture was diluted with water and EtOAc. The organic layer was isolatedand concentrated. The residue was separated by column chromatography(hexanes/EtOAc, 50:50) to afford the N-Boc protected title compound (300mg); ¹H NMR (500 MHz, CD₃OD) δ 1.43 (s, 9H), 2.40 (m, 1H), 2.82 (m, 2H),3.02 (m, 2H), 3.20 & 3.28 (ABX, J=15 and 8 Hz, 2H), 6.91 (m, 2H), 7.20(m, 1H); ¹³C NMR (125 MHz, D₂O) δ 27.77, 31.62, 38.39, 42.81, 52.34,79.23, 123.70. The aqueous layer was concentrated and the residue waspurified by column chromatography (CH₂Cl₂/MeOH 80/20) to afford a smallamount of the title compound (40 mg, 3%).

The N-Boc protected title compound (100 mg) in CH₂Cl₂ (2 mL) was treatedwith TFA (1 mL). The reaction mixture was stirred for 30 minutes,concentrated and suspended in EtOH. The white solid was collected byfiltration to give the title compound (65 mg, 93%); ¹H NMR (500 MHz,D₂O) δ 2.43 (m, 1H), 2.83 (dd, J=7.8 & 15 Hz, 1H), 2.85 (m, 1H), 2.92(m, 2H), 3.03 & 3.10 (ABX, J=13 and 7 Hz, 2H), 6.83 (m, 1H), 6.88 (m,1H), 7.20 (m, 1H); ¹³C NMR (125 MHz, D₂O) δ 31.68, 35.81, 42.44, 52.32,125.18, 127.04, 127.54, 140.31; ES-MS 234 (M−1).

Method 2:

Steps 1 to 3 are the same as those in Method 1.

To a solution of the mesylate (obtained in step 3) in CH₂Cl₂ (2 mL) wasadded TFA (1 mL). The reaction was stirred for 1 hour, concentrated toafford the corresponding amine TFA salt; ¹H NMR (500 MHz, D₂O) δ 2.39(m, 1H), 2.85-3.08 (m, 4H), 3.03 (s, 3H), 4.13 (m, 1H), 4.22 (m, 1H),6.83 (m, 1H), 6.88 (m, 1H), 7.18 (m, 1H).

A solution of the amine TFA salt (from step 4) in H₂O (5 mL) was addedslowly to a refluxed solution of Na₂SO₃ (1.17 g, 9.28 mmol). Thereaction mixture was stirred for 3 hours, and the solvent was removed.Concentrated HCl (5 mL) was added to precipitate the inorganic saltwhich was removed by filtration. The filtrate was concentrated; and theresultant residue was purified by column chromatography usingCH₂Cl₂/MeOH 90:10 to 80:20 as eluant to afford the title compound in 37%yield; NMR and MS spectra are identical to those from method 1.

Preparation of 4-amino-1-(2-thienyl)-2-butanesulfonic acid (CompoundN84)

To a solution of 2-thienylmethanol (5 g, 43.80 mmol) in dry ether (80mL) was added a solution of PBr₃ (17.7 g, 65.51 mmol) in ether (20 mL).The reaction mixture was stirred at room temperature for 30 minutes,quenched carefully by adding H₂O. The organic layer was isolated, washedwith brine and dried over Na₂SO₄. The solvent was removed carefully andthe residual material was purified rapidly by flash chromatography(hexanes/Et₂O 90:10 as eluant) to give 2-thienylmethyl bromide (6.2 g,80%).

To a cold (−50° C.) solution of 1,3-propane sultone (1.6 g, 13 mmol) inTHF (70 mL) was added a solution of BuLi (2 M in THF, 6.5 mL, 13 mmol).After stirring for 30 min, 2-thienylmethyl bromide (obtained in step 1,2.3 g, 13 mmol) was added and the reaction mixture was stirred for 2hours, and during this period the temperature was controlled between−50° C. and −20° C. The reaction mixture was cooled to −78° C., quenchedwith 1N HCl, and diluted with EtOAc. The organic phase was separated,washed with 1N HCl and concentrated. The residue was diluted withtoluene (10 mL) and separated by column chromatography (hexanes/EtOAc70:30 as eluant) to give 1-(2-thienyl)-2,4-butane sultone (800 mg, 35%);¹H NMR (500 MHz, CDCl₃) δ 2.40 (m, 1H), 2.60 (m, 1H), 3.15 (m, 1H), 3.55(m, 2H), 4.38 (m 1H), 4.43 (m, 1H), 6.94 (m, 2H), 7.22 (m, 1H). ¹³C NMR(125 MHz, CDCl₃) δ 29.31, 29.54, 56.76, 67.00, 125.29, 126.95, 127.60,137.92. 1,1-bis(2-thienylmethyl)-1,3-propane sultone was isolated in 9%yield (380 mg).

A solution of 1-(2-thienyl)-2,4-butane sultone (obtained in step 2, 218mg, 1 mmol) in a cosolvent of THF and EtOH (20 mL, v/v=1:1) was addedslowly to ammonium hydroxide (28%-30%, 10 mL, 80 mmol). The reactionmixture was stirred at room temperature for 4 hours, and thenconcentrated. The resulting solid was suspended in EtOH, heated atreflux for 15 minutes and cooled to room temperature. The solid materialwas collected by filtration, washed with ether and dried, finishing thetitle compound (170 mg, 71%); ¹H NMR (500 MHz, D₂O) δ 1.84-2.00 (m, 2H),2.80-3.10 (m, 4H), 3.38 (m, 1H), 6.88 (m, 2H), 7.20 (m, 1H); ¹³C NMR(125 MHz, D₂O) δ 27.05, 30.28, 37.78, 59.55, 125.16, 126.96, 127.54;ES-MS 234 (M−1).

Preparation of4-amino-1-(2-thienyl)-2-(2-thienylmethyl)-2-butanesulfonic acid(Compound N85)

To a solution of 1,1-bis(2-thienylmethyl)-1,3-propane sultone (obtainedin step 2 in the preparation of Compound N84,150 mg, 0.48 mmol) in acosolvent of THF and EtOH (10 mL, v/v=1:1) was added slowly an aqueoussolution of ammonium hydroxide (28%, 5 mL, 38 mmol). The reactionmixture was stirred at room temperature for 8 hours, and thenconcentrated. The resulting solid was suspended in EtOH, heated atreflux for 15 minutes and cooled to room temperature. The solid wascollected by filtration, washed with ether then dried, providing thetitle compound (140 mg, 88%); ¹H NMR (500 MHz, DMSO-d₆) δ1.78 (t, J=7.0Hz, 2H), 3.07-3.10 (m, 4H), 3.28 (s, 2H), 6.87 (m, 2H), 6.92 (m, 2H),7.30 (m, 2H), 7.60 (bs, 2H); ¹³C NMR (125 MHz, DMSO-d₆)

31.88, 34.96, 36.31, 60.74, 125.34, 127.37, 128.88, 139.60; ES-MS 330(M−1).

Preparation of 4-(tert-butylamino)-1-(thien-2-yl)butane-2-sulfonic acid(Compound N86)

To a solution of 1-(2-thienylmethyl)-1,3-propane sultone (obtained instep 2 from the preparation of Compound N84, 218 mg, 1 mmol) in THF (5mL) was added t-butylamine (1 mL). The reaction was stirred at roomtemperature for 15 hours, and then concentrated. The resulting solid wassuspended in EtOH (10 mL), heated at reflux for 1 hour and cooled toroom temperature. The solid was collected by filtration, washed withether then dried, giving the title compound (142 mg, 63%); ¹H NMR (500MHz, D₂O) δ 1.12 (s, 9H), 1.82 (m, 1H), 1.95 (m, 1H), 2.70 (m, 1H), 2.96(m, 2H), 3.07 (m, 1H), 3.40 (m, 1H), 6.90 (m, 2H), 7.21 (m, 1H). ¹³C NMR(125 MHz, D₂O) δ 24.92, 26.50, 30.25, 39.46, 57.16, 59.56, 125.27,126.99, 127.63; ES-MS 290 (M−1).

Preparation of 4-[2-(2-thien-2-ylpropyl)amino]-2-butanesulfonic acid(Compound N87)

To a stirred solution of 2-(2-thienyl)-2-propylamine (400 mg, 2.84 mmol)in THF (9 mL) was added 2,4-butane sultone (386 mg, 2.84 mmol). Thereaction mixture was stirred at reflux for 15 h, and then cooled to roomtemperature. The solid was collected by filtration, suspended in EtOH (5mL) and stirred at reflux for 1 hour. The suspension was then cooled toroom temperature. The solid was collected by filtration, washed withethanol and dried under high vacuum to afford the title compound (450mg, 57%); ¹H NMR (500 MHz, DMSO-d₆) δ 1.06 (d, J=6.8 Hz, 3H), 1.62 (m,1H), 1.71 (s, 6H), 1.95 (m, 1H), 2.75 (m, 1H), 2.85 (t, J=8.0 Hz, 2H),6.99 (m, 1H), 7.18 (m, 1H), 7.42 (m, 1H); ¹³C NMR (125 MHz, DMSO-d₆) δ14.65, 25.83, 25.87, 28.41, 40.20, 53.28, 59.50, 127.68, 127.95, 142.27;ES-MS 276 (M−1).

Preparation of4-[2-(2-thien-2-ylpropyl)amino]-1-(2-thienyl)-2-butanesulfonic acid(Compound N88)

To a stirred solution of 2-(2-thien-2-ylpropyl)amine (110 mg, 1.3 mmol)in pinacolne (9 mL) was added 1-(thien-2-ylmethyl)-1,3-propane sultone(obtained from step 2 in the preparation of Compound N84, 218 mg, 1.0mmol). The reaction mixture was stirred at reflux for 24 hours thencooled to room temperature. The solid was collected by filtration,suspended in EtOH (5 mL) and stirred at reflux for 1 hour. Thesuspension was then cooled to room temperature. The solid was collectedby filtration, washed with ethanol and dried under high vacuum to affordthe title compound (280 mg, 78%); ¹H NMR (500 MHz, DMSO-d₆) δ 1.68 (s,6H), 1.82 (m, 2H), 2.50-2.88 (m, 4H), 6.82 (m, 1H), 6.92 (m, 1H), 7.12(m, 1H), 7.26 (m, 1H), 7.34 (m, 1H), 7.65 (m, 1H), 9.00 (bs, 1H), 9.26(bs, 1H). ¹³C NMR (125 MHz, DMSO-d₆) δ 26.31, 26.56, 26.85, 31.24,58.41, 60.14, 125.06, 126.64, 127.74, 127.80, 127.92, 128.24, 142.11,144.00; ES-MS 358 (M−1).

Preparation of 3-(1-adamantyl)-3-aminopropane-1-sulfonic acid (CompoundQX)

A solution of 1-adamantanemethanol (10 g, 60 mmol) in dichloromethane(90 mL) was added to a suspension of pyridinium chlorochromate (18.5 g,86 mmol) in dichloromethane (120 mL) at room temperature over 15minutes. The reaction mixture was stirred at room temperature for 90minutes, followed by addition of heptane (450 mL). The mixture wasfiltered and the filtrate was concentrated. The residue was dissolved inpentane (100 mL) and the mixture was washed with NaOH (0.2N, 3×50 mL)and with water, dried over MgSO₄ and concentrated to giveadamantane-1-carbaldehyde as a white solid (7.2 g, 73%); ¹H NMR (CDCl₃,500 MHz) δ ppm 9.32 (s, 1H), 2.08-2.03 (m, 3H), 1.92-1.91 (m, 3H),1.80-L⁷⁵ (m, 3H), 1.73-1.68 (m, 6H).

Malonic acid (4.6 g, 43.8 mmol), ammonium acetate (7.0 g, 91 mmol) andadamantane-1-carbaldehyde from step 1 (7.2 g, 43.8 mmol) were refluxedin ethanol (11 mL) for 4 hours. The reaction mixture was cooled andfiltered. The solution was concentrated and the residue was distributedbetween dichloromethane and water. A precipitate, formed at this point,was collected by filtration. The water phase was freeze-dried; and thesolid material was combined with the solid obtained by filtration,giving 3-(1-adamantyl)-3-aminopropanoic acid as a white solid (6.82 g,70%); ¹H NMR (CD₃OD, 500 MHz) δ ppm 2.96 (dd, 1H, J=2.9 and 11.2 Hz),2.48 (dd, 1H, J=2.9 and 16.6 Hz), 2.23 (dd, 1H, J=11.7 and 16.6 Hz),2.04 (bs, 3H), 1.80-1.69 (m, 6H), 1.61 (m, 6H).

Borane (1 M solution in THF, 80.4 mL, 80.4 mmol) was added to a solutionof 3-(1-adamantyl)-3-aminopropanoic acid (from step 2, 5.98 g, 26.8mmol) in dry THF (25 mL). The reaction mixture was heated at reflux andstirred overnight. The reaction was quenched with a few drops ofmethanol then co-evaporated twice with methanol. The residue was treatedwith methanol (5 mL) and conc. HCl (5 mL) at reflux for 30 minutes. Thesolvent was evaporated and the residual material was dissolved in aminimum amount of methanol then precipitated with ether, collected byfiltration and dried under vacuum, to give3-(1-adamantyl)-3-aminopropan-1-ol as a white solid (4.77 g, 72%); ¹HNMR (CD₃OD, 500 MHz) δ ppm 3.86-3.82 (m, 1H), 3.75-3.71 (m, 1H), 2.91(d, 1H, J=10.7 Hz), 2.04 (bs, 3H), 1.98-1.94 (m, 1H), 1.80-1.78 (m, 3H),1.72-1.70 (m, 3H), 1.67-1.58 (m, 7H).

The alcohol from step 3 (4.77 g, 19.4 mmol) was refluxed in hydrobromicacid (47%, 45 mL) overnight. Solvent was removed under reduced pressureand the solid was dried under vacuum, providing1-(1-adamantyl)-3-bromopropan-1-amine as a white solid (6.04 g, 88%); ¹HNMR (CD₃OD, 500 MHz) δ ppm 3.70-3.66 (m, 1H), 3.55-3.49 (m, 1H), 2.93(dd, 1H, J=2.4 and 9.5 Hz), 2.39-2.32 (m, 1H), 2.06 (bs, 3H), 2.00-1.93(m, 1H), 1.81-1.78 (m, 3H), 1.73-1.70 (m, 3H), 1.62 (m, 6H).

1-(1-Adamantyl)-3-bromopropan-1-amine from step 4 (6.04 g, 17.1 mmol)and sodium sulfite (3.23 g, 25.7 mmol) were stirred in water at refluxovernight. The solid was collected by filtration and dried. It was thentreated with concentrated HCl (15 mL) and warmed. After 15 minutes, thesolution was cooled to room temperature and the solid was collected byfiltration, washed with a minimum amount of water, washed with ether anddried under vacuum, giving the title compound as a white solid (2.75 g,59%); ¹H NMR (DMSO, 500 MHz) δ ppm 7.83 (bs, 3H), 2.81-2.78 (m, 1H),2.72-2.70 (m, 1H), 2.06-2.02 (m, 1H), 2.01-1.97 (m, 3H), 1.68-1.49 (m,13H); ¹³C NMR (DMSO, 500 MHz) δ ppm 107.0, 61.0, 50.1, 37.7, 36.8, 35.3,28.1, 23.7; ES-MS 272 (M−1).

Preparation of 2-(2-adamantyl)-3-aminopropane-1-sulfonic acid (CompoundSX)

To a solution of diisopropylamine (5.0 mL, 0.036 mol) in THF (30 mL) wasadded at −78° C. n-BuLi (21 mL, 1.6 M, 0.035 mol). The solution wasstirred at −78° C. for 30 minutes, then warmed to 0° C. and stirred foran additional 30 minutes. The solution was then cooled to −78° C. andtreated with a solution of 2-adamantaneacetonitrile (4.5 g, 0.015 mol)in THF (10 mL). The mixture was stirred at −78° C. for 30 minutes, andwarmed to room temperature for 30 minutes. The mixture was then cooledto −78° C. and treated with diethylcarbonate (2.0 mL, 0.016 mol). Thereaction mixture was stirred at −78° C. for 3 hours. The mixture wasquenched with a saturated aqueous solution of NH₄Cl (10 mL), followed byaddition of water. The aqueous phase was extracted with ether and thecombined organic phase was washed with 5% HCl, and dried over MgSO₄.Hexane was added and the mixture was filtered. The filtrate was purifiedby Biotage (5% EtOAc in hexanes) to furnish ethylα-cyano-adamantane-2-acetate (2.74 g, 74%) containing about 20% ofstarting material. The material was used in the following reactionwithout further purification.

To a solution of crude ethyl ester (from step 1, 1.29 g, 5.22 mmol) inTHF (4 mL) was added LAH 2.0 M (8.0 mL, 16 mmol) slowly. The mixture washeated at reflux overnight, quenched with a minimum amount of 20% KOH inwater. The mixture was filtered through Celite™. The filtrate wasevaporated under vacuum; and the residue was dissolved indichloromethane (15 mL). To this dichloromethane solution was added(BOC)₂O (1.5 g) and the mixture was stirred overnight. Solvent wasremoved by evaporation and the resulting residue was purified onBiotage, affording 2-(2-adamantyl)-N-Boc-3-amino-1-propanol (425 mg,26%).

2-(2-Adamantyl)-N-Boc-3-amino-1-propanol (425 mg from step 2, 1.38 mmol)was treated with aqueous HBr 48% (20 mL) and heated at reflux overnight.Water was evaporated and solid was triturated with ether to afford2-(2-adamantyl)-3-bromo-1-propylamine HBr salt (486 mg, 100%) as beigesolid.

2-(2-Adamantyl)-3-bromo-1-propylamine HBr salt (486 mg from step 3, 1.38mmol) and sodium sulfite (260 mg, 2.06 mmol) in degassed water (3 mL)were heated at reflux overnight. The solid was filtered and washed withwater (20 mL). The solid was transferred in methanol (20 mL), and themixture was refluxed for 1 hour. The solid material was collected byfiltration, providing the title compound (105 mg, 28%); ¹H NMR (500 MHz,DMSO-d6) δ 3.06 (dd, J=1.9 Hz, J=12.7 Hz, 1H), 2.73 (m, 2H), 2.60 (m,1H), 2.31 (m, 1H), 1.84-1.79 (m, 8H), 1.68-1.62 (m, 4H), 1.45 (m, 3H);ES-MS 272 (M−1).

Preparation of 3-(2-adamantyl)-3-aminopropane-1-sulfonic acid (CompoundSY)

A mixture of adamantane-2-carbaldehyde (9.6 g, 0.058 mol), ammoniumacetate (9.4 g, 0.12 mol) and malonic acid (6.0 g, 0.058 mol) in ethanol(30 mL) was heated at reflux overnight. The mixture was cooled to roomtemperature; and the solid was removed by filtration. The filtrate waspoured into a mixture of dichloromethane/water (150 mL/150 mL). Thesolid thus formed was collected by filtration and dried under vacuum,giving 3-amino-3-(2-adamantyl)propanoic acid (4.6 g, 35%)

To a solution of the above obtained acid (2.0 g, 0.0090 mol) in dry THF(10 mL) was added BH₃-THF 1 M (27 mL, 0.0027 mol). The mixture washeated at reflux overnight. The reaction mixture was cooled to roomtemperature, and quenched carefully with methanol (1 mL). Evaporation ofthe solvent left a residue which was then treated with methanol (20 mL)and concentrated HCl (20 mL) at reflux for 30 minutes. All volatileswere evaporated and the solid residue was dissolved in methanol (100 mL)and precipitated with ether (150 mL). The solid was collected byfiltration, providing the 3-amino-3-(2-adamantyl)-1-propanol as ahydrochloride salt (1.6 g, 74%).

A suspension of the amino alcohol (820 mg from step 2, 3.34 mmol) inaqueous HBr solution (48%, 25 mL) was heated at reflux overnight. Anadditional amount of aqueous HBr (48%, 25 mL) was added and the mixturewas refluxed for additional 24 hours. Evaporation of the solvent gave3-bromo-1-(2-adamantyl)-propylamine as HBr salt (1.27 g, 100%).

The product of step 3 (1.27 g, 3.59 mmol) and Na₂SO₃ (680 mg, 5.40 mmol)in degassed water (10 mL) was heated at reflux overnight. The mixturewas cooled to room temperature; and the solid was collected byfiltration and washed with a small amount of water (5-10 mL). Theresulting solid was treated with ethanol (10 mL) at reflux for 30minutes. The solid was collected by filtration and dried in vacuo,giving the title compound (658 mg, 67%) as a white solid; ¹H NMR (500MHz, CD₃OD), δ 3.69 (m, 1H), 2.87 (t, 2H, J=6.3 Hz), 2.22 (m, 1H),1.93-1.73 (m, 14H), 1.57 (m, 2H); ES-MS 272 (M−1).

Preparation of 2-(1-adamantyl)-3-aminopropane-1-sulfonic acid (CompoundSZ)

A solution of ethyl methanesulfonate (1.21 mL, 11.7 mmol) in THF (10 mL)was cooled to −78° C. To this cold solution was added LiHMDS 1M solutionin THF (12.5 mL, 12.5 mmol) dropwise. The yellow solution was stirredfor 30 minutes, and then transferred via cannula to a second flaskcontaining a solution of adamantyl bromomethyl ketone (2.00 g, 7.80mmol) in THF (40 mL) at −78° C. The mixture was stirred at −78° C. for1.5 hours, warmed slowly to −45° C., and further stirred for 2.5 hours.The reaction was quenched by adding a saturated aqueous solution ofNH₄Cl (100 mL). The mixture was extracted with AcOEt (3×100 mL), washedwith brine, dried over MgSO₄, and concentrated under reduced pressure togive 2-(1-admantyl)-1,3-prop-1-ene sultone as a yellow solid (1.93 g,97%).

Palladium on activated carbon (10%, 0.198 g) was added to a solution ofthe above obtained sultone (0.66 g, 2.59 mmol) in MeOH (60 mL) in aPyrex™ high pressure bottle, and hydrogenated at 40 psi for 48 hours.The reaction mixture was filtered through Celite™ and the filtrate wasconcentrated to dryness, providing 2-(1-admantyl)-1,3-propane sultone asa white solid (0.65 g, 98%).

2-(1-adamantyl)-1,3-propane sultone (0.620 g from step 2, 2.42 mmol) wasdissolved in DMF (10 mL). NaN₃ (0.188 g, 2.90 mmol) was added and thesolution was heated at 130° C. overnight. Solvent was evaporated and abeige solid was obtained. The solid was dissolved in methanol, and thesolution was passed through Amberlite™ IR-120(plus) ion-exchange resin(10 g, pre-washed with water and then MeOH). Evaporation of the solventgave a yellow viscous liquid which became beige solid under vacuum (0.43g, 76%).

Palladium on activated carbon (10%, 0.150 g) was added to a solution ofthe sulfonic acid derivative from step 3 (0.4176 g, 1.79 mmol) in asolvent mixture of methanol (36 mL) and acetic acid (4 mL) in a Pyrex™high pressure bottle. The mixture was hydrogenated at 30 psi overnight.The reaction mixture was filtered through Celite™ and the filtrate wasconcentrated to dryness, giving the title compound as a white solid(0.210 g, 57%); ¹H NMR (DMSO, 500 MHz) δ ppm 7.81 (large s, 3H), 3.11(d, 1H, J=12.7 Hz), 2.90 (d, 1H, J=13.7 Hz), 2.70 (dd, 1H, J=10.1 Hz,J=12.4 Hz), 1.9 (large s, 3H), 1.67-1.59 (m, 7H), 1.49-1.31 (m, 6H). 1 His partially hidden with DMSO signal.

Preparation of 2-[(benzylamino)methyl]-3,3-dimethylbutane-1-sulfonicacid (Compound N42)

To a solution of 4-tert-butyl-1,2-oxathiolane 2,2-dioxide (167 mg, 0.94mmol) in dimethylformamide (2 mL) was added benzylamine (0.153 mL). Thesolution was stirred at 130° C. for 24 hours. The reaction mixture wascooled to room temperature and triturated with ethanol. The solidmaterial was collected by filtration and washed successively withethanol (2×20 mL) and with acetone (2×20 mL), giving the title compound(205 mg, 77%); ¹H NMR (CD₃OD, 500 MHz) δ ppm 7.24 (m, 5H), 4.21 (q, 2H,J=13 & 12.5 Hz), 3.43 (d, 1H, J=13 Hz), 3.23 (d, 1H, J=14 Hz), 3.09 (m,1H), 2.87 (dd, 1H, J=9.5, J=10 Hz), 2.05 (br t, 1H, J=9.5 Hz), 1.02 (s,9H); ES-MS 284 (M−1).

Preparation of 3-(9H-fluoren-9-ylamino)propane-1-sulfonic acid (CompoundN44)

9-Aminofluorene hydrochloride (1.02 g, 4.69 mmol) was treated with asaturated solution of K₂CO₃ (100 mL) and the mixture was extracted withEtOAc (3×75 mL). The organic extracts were combined, dried over Na₂SO₄,concentrated to furnish a residue, which was azeotroped with toluene toyield 9-aminofluorene (0.831 g, 4.59 mmol). The free amine was dissolvedin 25% toluene/acetonitrile (10 mL). To the resulting solution was added1,3-propane sultone (532 mg, 4.36 mmol). The reaction mixture wasstirred at reflux for 4 h, and cooled to room temperature. The solidmaterial was collected by filtration, washed with acetone, and thentreated with ethanol at reflux for 1 hours. After the mixture was cooledto room temperature, the solid was collected by filtration, washed withacetone, and dried in vacuo, providing the title compound (808 mg, 74%);¹H NMR (DMSO, 500 MHz) δ ppm 9.96 (large s, 2H), 7.96 (d, 2H, J=7.3 Hz),7.88 (d, 2H, J=7.8 Hz), 7.56 (t, 2H, J=7.3 Hz), 7.45 (t, 2H, J=7.3 Hz),5.66 (s, 1H), 2.98 (s, 1H), 2.64 (t, 2H, J=5.9 Hz), 1.96 (t, 2H, J=5.9Hz). ¹³C NMR (CD₃OD, 125 MHz) δ ppm 141.02, 137.63, 130.10, 128.12,126.15, 120.80, 59.44, 49.64, 43.43, 21.82.

3-[(4,7-dimethoxyindan-1-yl)amino]propane-1-sulfonic acid (Compound N51)

To a stirred solution of 4,7-dimethoxy-1-indanone (515 mg, 2.67 mmol) inethanol (10 mL) was added a suspension of hydroxylamine hydrochloride(447 mg, 6.43 mmol) in ethanol/water (1 mL/1 mL), followed by additionof a suspension of sodium acetate (527 mg, 6.43 mmol) in ethanol/water(1 mL/1 mL). The reaction mixture was heated at reflux for 3.5 hours.Water was added and the white solid thus formed was collected byfiltration, furnishing 4,7-dimethoxyindan-1-one oxime (463 mg, 84%).

Palladium on activated carbon (10%, 100 mg) was added to a degassedsolution of the 4,7-dimethoxyindan-1-one oxime obtained from step 1 (463mg, 2.23 mmol) in a mixture of methanol (45 mL) and acetic acid (5 mL).The reaction mixture was stirred under hydrogen atmosphere (1 Atm)overnight. The reaction mixture was flushed with nitrogen and filteredthrough Celite™. The filtrate was concentrated and the residual materialwas azeotroped twice with toluene to yield 1-amino-4,7-dimethoxyindan(quantitative).

To a solution of 1-amino-4,7-dimethoxyindan (2.23 mmol) in a 25%acetonitrile/toluene solution (20 mL) was added 1,3-propane sultone (259mg, 2.12 mmol). The reaction mixture was heated at reflux for 4 hours,and then cooled to room temperature. The mixture was concentrated to abeige solid; and the solid material was dissolved in ethanol by heating.The hot mixture was cooled to room temperature and left standingovernight. The crystalline solid thus formed was collected by filtrationand dried in vacuo, providing the title compound as a white solid (223mg, 32%); ¹H NMR (DMSO, 500 MHz) δ ppm 8.86 (bs, 1H), 8.66 (bs, 1H),6.95 (d, 1H, J=8.8 Hz), 6.85 (d, 1H, J=8.8 Hz), 4.73 (m, 1H), 3.78 (s,3H), 3.75 (s, 3H), 3.05 (m, 2H), 3.00-2.94 (m, 1H), 2.79-2.74 (m, 1H),2.62-2.60 (m, 2H), 2.39-2.34 (m, 1H), 2.19-2.15 (m, 1H), 1.98 (m, 2H);ES-MS 314 (M−1).

Preparation of 3-[(5,6-dimethoxyindan-1-yl)amino]propane-1-sulfonic acid(Compound N53)

To a stirred solution of 5,6-dimethoxy-1-indanone (565 mg, 2.94 mmol) inethanol (10 mL) was added a suspension of hydroxylamine hydrochloride(490 mg, 7.05 mmol) in ethanol/water (1 mL/1 mL), followed by additionof a suspension of sodium acetate (578 mg, 7.05 mmol) in ethanol/water(1 mL/1 mL). The reaction mixture was heated at reflux for 3.5 hours.Water was then added and the white solid was collected by filtration anddried in vacuo to provide 5,6-dimethoxy-1-indanone oxime (488 mg, 80%).

Palladium on activated carbon (10%, 80 mg) was added to a solution ofthe oxime obtained from step 1 (285 mg, 1.37 mmol) in methanol (36 mL)and acetic acid (4 mL). The reaction mixture was stirred under hydrogenatmosphere (1 Atm) overnight. The reaction mixture was flushed withnitrogen and filtered through Celite™. The filtrate was concentrated andthe residue was azeotroped twice with toluene to yield1-amino-5,6-dimethoxy-1-indan as a light yellow solid (quantitative),containing a small amount of solvent.

To a solution of 1-amino-5,6-dimethoxy-1-indan (1.37 mmol) in a 25%toluene/acetonitrile solution (10 mL) was added 1,3-propane sultone (159mg, 1.30 mmol). The reaction mixture was stirred at reflux for 4 hours.The white solid formed was collected by filtration, washed with acetone,and then treated in ethanol at refluxed for 1 hour. The ethanolicmixture was cooled to room temperature; and the solid material wascollected by filtration, washed with acetone and dried in vacuo tofurnish the title compound (191 mg, 44%); ¹H NMR (DMSO, 500 MHz) δ ppm8.87 (large s, 2H), 7.17 (s, 1H), 6.94 (s, 1H), 4.65-4.64 (m, 1H), 3.76(s, 6H), 3.14-2.98 (m, 3H), 2.83-2.77 (m, 1H), 2.68-2.65 (m, 2H),2.44-2.37 (m, 1H), 2.15-2.11 (m, 2H); ES-MS: 314 (M−1).

Preparation of 3-[5-methoxyindan-1-yl)amino]propane-1-sulfonic acid(Compound N56)

To a stirred solution of 5-methoxy-1-indanone (1.06 g, 6.5 mmol) inethanol (20 mL) was added a suspension of hydroxylamine hydrochloride(1.08 g, 15.6 mmol) in ethanol/water (2 mL/2 mL), followed by theaddition of a suspension of sodium acetate (1.28 g, 15.6 mmol) inethanol/water (2 mL/2 mL). The reaction mixture was heated at reflux for3.5 hours. Water was added and the beige solid was collected byfiltration, giving 5-methoxyindan-1-one oxime (1 g, 87%).

Palladium on activated carbon (10%, 100 mg) was added to a solution of5-methoxyindan-1-one oxime from step 1 (1 g, 5.64 mmol) in methanol (90mL) and acetic acid (10 mL). The reaction mixture was stirred under anhydrogen atmosphere overnight, then was flushed with nitrogen andfiltered through Celite™. The filtrate was concentrated and the residualmaterial was azeotroped twice with toluene to give1-amino-5-methoxyindan (quantitative).

To a solution of 1-amino-5-methoxy-1-indan (5.64 mmol) in a 25%acetonitrile/toluene solution (50 mL) was added 1,3-propane sultone (654mg, 5.36 mmol). The reaction mixture was stirred at reflux for 4 hours.The white solid formed was collected by filtration, transferred inethanol and heated at reflux for 1 hour. After cooling to roomtemperature, the solid was collected by filtration and dried in vacuo toprovide the title compound as a white solid (985 mg, 64%); ¹H NMR (DMSO,500 MHz) δ ppm 9.05 (bs, 1H), 8.86 (bs, 1H), 7.46 (d, 1H, J=8.8 Hz),6.92 (d, 1H, J=2 Hz), 6.86 (dd, 1H, J=2.4 and 8.5 Hz), 4.65-4.63 (m,1H), 3.76 (s, 3H), 3.11 (t, 2H, J=6.8 Hz), 3.08-3.03 (m, 1H), 2.87-2.81(m, 1H), 2.69-2.64 (m, 2H), 2.44-2.37 (m, 1H), 2.17-2.11 (m, 1H),2.00-1.95 (m, 2H); ¹³C NMR (DMSO, 500 MHz) δ ppm 161.3, 147.3, 130.2,127.2, 113.8, 110.6, 61.7, 56.0, 50.0, 45.4, 30.7, 29.1, 22.5; ES-MS 284(M−1).

Preparation of 3-[(5-fluoroindan-1-yl)amino]propane-1-sulfonic acid(Compound N58)

To a stirred solution of 5-fluoro-1-indanone (1 g, 6.7 mmol) in ethanol(20 mL) was added a suspension of hydroxylamine hydrochloride (1.11 g,16 mmol) in ethanol/water (2 mL/2 mL), followed by the addition of asuspension of sodium acetate (1.31 g, 16 mmol) in ethanol/water (2 mL/2mL). The reaction mixture was heated at reflux for 3.5 h. Water wasadded and the white solid was collected by filtration, giving5-fluoroindan-1-one oxime (1.1 g, 99%).

Palladium on activated carbon (10%, 200 mg) was added to a solutionflushed with nitrogen of the 5-fluoroindan-1-one oxime obtained in step1 (1.1 g, 6.6 mmol) in methanol (90 mL) and acetic acid (10 mL). Thereaction mixture was and stirred under an hydrogen atmosphere overnight.The reaction mixture was flushed with nitrogen and filtered throughCelite™. The filtrate was concentrated and residual material wasazeotroped twice with toluene to yield 1-amino-5-flluoroindan(quantitative).

To a solution of 1-amino-5-fluoroindan (6.6 mmol) in a 25%acetonitrile/toluene solution (50 mL) was added 1,3-propane sultone (766mg, 6.3 mmol). The mixture was heated at reflux for 4 hours. The whitesolid material thus formed was collected by filtration, transferred inethanol and heated at reflux for 1 hour. After cooling to roomtemperature, the solid was collected by filtration and dried in vacuo tofurnish the title compound as a white solid (1.16 g, 68%); ¹H NMR (DMSO,500 MHz) δ ppm 9.16 (bs, 1H), 8.96 (bs, 1H), 7.60 (dd, 1H, J=8.3 and 5.4Hz), 7.21 (dd, 1H, J=2 and 9 Hz), 7.17-7.13 (m, 1H), 4.73-4.70 (m, 1H),3.18-3.13 (m, 2H), 3.11-3.06 (m, 1H), 2.92-2.86 (m, 1H), 2.71-2.65 (m,2H), 2.47-2.41 (m, 1H), 2.21-2.14 (m, 1H), 2.03-1.96 (m, 2H); ES-MS 272(M−1).

Preparation of 3-[(2-methylindan-1-yl)amino]propane-1-sulfonic acid(Compound N59)

To a stirred solution of 2-methyl-1-indanone (1 g, 6.8 mmol) in ethanol(20 mL) was added a suspension of hydroxylamine hydrochloride (1.14 g,16 mmol) in ethanol/water (2 mL/2 mL), followed by addition of asuspension of sodium acetate (1.34 g, 16 mmol) in ethanol/water (2 mL/2mL). The reaction mixture was heated at reflux for 5 h, cooled to roomtemperature, and poured into water. The mixture was extracted twice withAcOEt. The extracts were combined, washed with brine, dried over sodiumsulfate, and concentrated to dryness, giving 2-methylindan-1-one oximeas colorless oil (quantitative).

Palladium on activated carbon (10%, 200 mg) was added to a solution of2-methylindan-1-one oxime obtained from step 1 (1.1 g, 6.6 mmol) inmethanol (90 mL) and acetic acid (10 mL). The reaction mixture wasstirred under hydrogen atmosphere overnight, then flushed with nitrogenand filtered through Celite™. The filtrate was concentrated and theresidue was azeotroped twice with toluene, providing1-amino-2-methylindan (quantitative).

To a solution of 1-amino-2-methylindan (6.8 mmol) in a 25%acetonitrile/toluene solution (50 mL) was added 1,3-propane sultone (794mg, 6.5 mmol). The reaction mixture was stirred at reflux for 4 hours,and then cooled to room temperature. The white solid was collected byfiltration, transferred in ethanol and heated at reflux for 30 min.After the mixture was cooled to room temperature, and the solid formedwas collected by filtration and dried in vacuo to give the titlecompound as a mixture of isomers (267 mg, 15%); ¹H NMR of the majorisomer (DMSO, 500 MHz) δ ppm 8.95 (bs, 2H), 7.51 (d, 1H, J=7.3 Hz),7.38-7.33 (m, 2H), 7.29-7.26 (m, 1H), 4.54-4.53 (m, 1H), 3.18-3.17 (m,1H), 3.10-3.08 (m, 1H), 2.99-2.95 (m, 1H), 2.82-2.75 (m, 2H), 2.70-2.67(m, 2H), 2.04-1.99 (m, 2H), 1.17 (d, 3H, J=6.8 Hz); ES-MS 268 (M−1).

Preparation of3-[(4-methoxy-2,3-dihydro-1H-inden-1-yl)amino]propane-1-sulfonic acid(Compound N61)

To a stirred solution of 4-methoxy-1-indanone (1 g, 6.5 mmol) in ethanol(20 mL) was added a suspension of hydroxylamine hydrochloride (1.08 g,16 mmol) in ethanol/water (2 mL/2 mL), followed by the addition of asuspension of sodium acetate (1.28 g, 16 mmol) in ethanol/water (2 mL/2mL). The reaction mixture was heated at reflux for 4 hours, cooled toroom temperature, followed by addition of water. The solid material wascollected by filtration, affording 4-methoxy-1-indanone oxime (1.1 g,quantitative).

Palladium on activated carbon (10%, 200 mg) was added to a solution ofthe 4-methoxyindan-1-one oxime obtained from step 1 (1.1 g, 6.5 mmol) inmethanol (90 mL) and acetic acid (10 mL). The reaction mixture wasstirred under hydrogen atmosphere overnight. The reaction mixture wasflushed with nitrogen and filtered through Celite™. The filtrate wasconcentrated and the residue was azeotroped twice with toluene to yield4-methoxyindan-1-amine (quantitative).

To a solution of 4-methoxyindan-1-amine (6.5 mmol) in a 25%acetonitrile/toluene solution (50 mL) was added 1,3-propane sultone (754mg, 6.2 mmol). The reaction mixture was stirred at reflux for 4 hours.The white solid was collected by filtration, transferred in ethanol andheated at reflux for 30 minutes. After cooling to room temperature, thesolid material was collected by filtration and dried in vacuo,furnishing the title compound as a white solid (1.08 g, 59%); ¹H NMR(DMSO, 500 MHz) δ ppm 9.20 (bs, 1H), 8.94 (bs, 1H), 7.31 (t, 1H, J=8.3Hz), 7.15 (d, 1H, J=7.3 Hz), 6.99 (d, 1H, J=8.3 Hz), 4.75-4.73 (m, 1H),3.81 (s, 3H), 3.12-3.11 (m, 2H), 2.98-2.92 (m, 1H), 2.81-2.75 (m, 1H),2.68-2.66 (m, 2H), 2.45-2.37 (m, 1H), 2.16-2.10 (m, 1H), 2.00-1.95 (m,2H). ¹³C NMR (DMSO, 500 MHz) δ ppm 156.5, 139.9, 132.6, 129.4, 118.0,111.8, 62.4, 55.9, 50.0, 45.5, 28.4, 27.5, 22.5; ES-MS 284 (M−1).

Preparation of 3-[(6-methoxyinden-1-yl)amino]propane-1-sulfonic acid(Compound N62)

To a stirred solution of 6-methoxyindan-1-one 1 (1.00 g, 6.16 mmol) inethanol (20 mL) was added to a suspension of hydroxylamine hydrochloride(1.03 g, 14.8 mmol) in ethanol/water (2 mL/2 mL), followed by theaddition of a suspension of sodium acetate (1.28 g, 15.6 mmol) inethanol/water (4 mL/4 mL). The reaction mixture was heated at reflux for3 hours. Water was added to the reaction mixture; and the white solidthus formed was collected by filtration, giving 6-methoxyindan-1-oneoxime (0.81 g, 74%).

Palladium on activated carbon (10%, 180 mg) was added to a solution ofthe oxime (obtained from step 1, 0.810 g, 4.57 mmol) in methanol (54 mL)and acetic acid (6 mL). The reaction mixture was stirred under anhydrogen atmosphere overnight, and then flushed with nitrogen andfiltered through Celite™. The filtrate was concentrated and the residuewas azeotroped twice with toluene to yield 6-methoxyindan-1-amine(quantitative) as beige solid, containing a small amount of solventwhich was used in next step without purification.

To a solution of 6-methoxyindan-1-amine (4.57 mmol) in a 25%toluene/acetonitrile solution (20 mL) was added 1,3-propane sultone (714mg, 5.85 mmol). The reaction mixture was heated at reflux for 4 hours.The white solid thus formed was collected by filtration, washed withacetone, transferred in ethanol and heated at reflux for 1 hour, andthen cooled to room temperature. The solid was collected by filtration,washed with acetone and dried in vacuo, providing the title compound asa white solid (568 mg, 44%); ¹H NMR (CD₃OD, 500 MHz) δ ppm 7.25 (d, 1H,J=8.3 Hz), 7.15 (d, 1H, J=2.0 Hz), 6.95 (dd, 1H, J=2.4 Hz, J=8.8 Hz),4.75 (dd, 1H, J=4.4 Hz, J=7.8 Hz), 3.81 (s, 3H), 3.30-3.24 (m, 2H),3.13-3.07 (m, 1H), 2.97-2.88 (m, 3H), 2.61-2.53 (m, 1H), 2.27-2.21 (m,1H), 2.19-2.14 (m, 2H)¹³C NMR (CD₃OD, 125 MHz) δ ppm 161.00, 139.50,137.73, 127.29, 117.89, 111.22, 64.37, 56.20, 50.07, 46.24, 30.37,30.35, 23.24; ES-MS 284 (M−1).

3-[(4-methylindan-1-yl)amino]propane-1-sulfonic acid (Compound N68)

To a stirred solution of 4-methyl-1-indanone (1 g, 6.8 mmol) in ethanol(20 mL) was added a suspension of hydroxylamine hydrochloride (1.14 g,16 mmol) in ethanol/water (2 mL/2 mL), followed by the addition of asuspension of sodium acetate (1.35 g, 16 mmol) in ethanol/water (2 mL/2mL). The reaction mixture was heated at reflux for 4 hours. Water wasadded to the reaction mixture; and the white solid thus formed wascollected by filtration, giving 4-methyl-1-indanone oxime (1.1 g,quantitative).

Palladium on activated carbon (10%, 200 mg) was added to anitrogen-flushed solution of the 4-methoxyindan-1-one oxime (1.1 g, 6.8mmol) in methanol (90 mL) and acetic acid (10 mL). The reaction mixturewas stirred under hydrogen atmosphere overnight, flushed with nitrogenand filtered through Celite™. The filtrate was concentrated and theresidual material was azeotroped twice with toluene to yield4-methoxyindan-1-amine (quantitative).

To a solution of 4-methoxyindan-1-amine (6.8 mmol) in a 25%acetonitrile/toluene solution (50 mL) was added 1,3-propane sultone (794mg, 6.5 mmol). The reaction mixture was stirred at reflux for 4 hours.The white solid obtained was collected by filtration, transferred inethanol and heated at reflux for 30 minutes, and then cooled to roomtemperature. The solid was collected by filtration and dried in vacuo tofurnish the title compound as a white solid (395 mg, 25%); ¹H NMR (DMSO,500 MHz) δ ppm 9.19 (bs, 1H), 8.92 (bs, 1H), 7.39 (d, 1H, J=7.3 Hz),7.24-7.19 (m, 2H), 4.74 (m, 1H), 3.14 (m, 2H), 3.02-2.96 (m, 1H),2.84-2.78 (m, 1H), 2.68-2.66 (m, 2H), 2.44-2.38 (m, 1H), 2.25 (s, 3H),2.16-2.10 (m, 1H), 2.01-1.97 (m, 2H); ES-MS 268 (M−1).

(3R)-3-amino-5-methylhexane-1-sulfonic acid (Compound N73)

Di t-butyldicarbonate (9 g, 41 mmol) was added to a solution of leucinemethyl ester hydrochloride salt (5 g, 28 mmol) and triethylamine (7.7mL, 55 mmol) in dichloromethane (200 mL). The reaction mixture wasstirred at room temperature overnight, washed with citric acid (10%),saturated NaHCO₃, and brine subsequently, and dried over sodium sulfate.The solution was concentrated to give an oil residue. The residue waspurified by flash chromatography using EtOAc/hexane 30%, givingBoc-Leu-OMe as colorless oil (quantitative).

In a dry 3-neck 500 mL flask equipped with a low temperature thermometerwas added the above-obtained ester (27.5 mmol) and toluene (70 mL). Thesolution was cooled to −78° C. then Diisobutyl aluminum hydride 1Msolution in toluene (38.5 mL 38.5 mmol) was added dropwise using anaddition funnel so that the internal temperature was kept under −65° C.(over 2 h). The reaction mixture was stirred at −78° C. for anadditional 2 hours then it was quenched by slowly adding 10 mL of coldmethanol (−78° C.) also keeping the internal temperature below −65° C.The reaction mixture was slowly poured into 100 mL of ice-cold HCl 1Nsolution with stirring over 15 minutes. The aqueous mixture wasextracted with EtOAc (3×500 mL). The organic layers were combined,washed with brine, dried over sodium sulfate, and concentrated to givethe corresponding aldehyde as colorless oil (quantitative).

Ethyl(diethoxyphosphoryl)methanesulfonate (4.8 g, 18.6 mmol) was addedto a suspension of sodium hydride (334 mg, 14 mmol) in dry THF (100 mL)at 0° C. The reaction mixture was stirred at 0° C. for 15 minutes, thenthe aldehyde (2 g, 9.3 mmol) was added in one portion. The reactionmixture was stirred at 0° C. for 2 h, then at room temperature for 1hour. The mixture was poured into water, extracted with EtOAc. Thecombined extracts were washed with brine, dried over sodium sulfate, andconcentrated to give colorless oil. Purification of the oil residue byBiotage chromatography using EtOAc/hexane 50% gave ethyl(1E,3R)-3-[(tert-butoxycarbonyl)amino]-5-methylhex-1-ene-1-sulfonate ascolorless oil which crystallized on the pump (2.43 g, 81%).

Palladium on activated charcoal (10%, 200 mg) was added to a solution ofthe product from step 3 (1.89 g, 5.87 mmol) in ethanol (50 mL) under N₂atmosphere. The reaction mixture was stirred under hydrogen atmospherefor 1.5 hours. The mixture was flushed with nitrogen, passed throughCelite™ pad and washed with ethanol several times. Solvent was removedto give ethyl(3R)-3-[(tert-butoxycarbonyl)amino]-5-methylhexane-1-sulfonate ascolorless oil which solidified on standing in vacuo. (1.72 g, 91%)

The reaction product from step 4 (1.72 g, 5.3 mmol) in formic acid (15mL) and water (0.6 mL) was heated at 50° C. for 24 hours. The reactionmixture was concentrated and dried in vacuo. The gum-like material thusobtained was dissolved in a minimum amount of methanol and precipitatedwith ether. The solid was collected by filtration. The precipitationstep was done twice to yield the title compound as a white solid (0.571g, 55%); ¹H NMR (D₂O, 500 MHz) δ ppm 3.39-3.36 (m, 1H), 2.89 (t, 2H,J=7.3 Hz), 1.98-1.91 (m, 2H), 1.57-1.55 (m, 1H), 1.41-1.36 (m, 2H), 0.78(dd, 6H, J=3.4 and 7.2 Hz) ¹³C NMR (D₂O, 500 MHz) δ ppm 49.1, 46.9,41.0, 27.9, 23.9, 21.8, 21.3; ES-MS 194 (M−1).

Preparation of (1E,3R)-3-amino-5-methylhex-1-ene-1-sulfonic acid(Compound N74)

Ethyl(1E,3R)-3-[(tert-butoxycarbonyl)amino]-5-methylhex-1-ene-1-sulfonate(see step 3 in the preparation of NRM8588) (532 mg, 1.7 mmol) in formicacid (5 mL) and water (0.2 mL) was heated at 50° C. for 24 hours. Thereaction mixture was concentrated and dried in vacuo. The gum-likematerial thus obtained was dissolved in a minimum amount of methanol andprecipitated with ether. The solid was collected by filtration. Theprecipitation step was done twice to yield the title compound as a whitesolid (0.082 g, 26%); ¹H NMR (D₂O, 500 MHz) δ ppm 6.57 (d, 1H, J=15 Hz),6.30 (dd, 1H, J=8.2 and 15 Hz), 3.90-3.87 (m, 1H), 1.55-1.46 (m, 3H),0.80-0.77 (m, 6H). ES-MS 192 (M−1).

Preparation of2-{[1R)-2,3-dihydro-1H-inden-1-ylamino]methyl}-3,3-dimethylbutane-1-sulfonicacid (Compound N76)

To a solution of 4-tert-butyl-1,2-oxathiolane 2,2-dioxide (225 mg, 1.26mmol) in dimethylformamide (5 mL) was added excess of (R)-indanamine(410 mg). The solution was stirred at 130° C. for 48 hours. The reactionmixture was cooled to room temperature and the solvent was evaporatedunder reduced pressure. The residue was triturated with ethanol. Thesolid material was collected by filtration and washed with acetone (2×20mL) to give the title compound (295 mg, 75% yield); ¹H NMR (CD₃OD, 500MHz) δ ppm 7.43 (m, 1H), 7.16 (m, 3H), 4.25 (m, 1H), 3.14 (m, 1H), 2.00(m, 2H), 2.72 (m, 2H), 2.69 (m, 1H), 2.23 (m, 1H), 1.88 (m, 2H), 1.00(s, 9H); ES-MS 310 (M−1).

Preparation of 3-[(4,5-dimethoxyinadan-1-yl)amino)propane-1-sulfonicacid (Compound N77)

A mixture of 4,5-dimethoxy-1-indanone (1.06 g, 5.5 mmol), hydroxylaminehydrochloride (0.92 g), and sodium acetate (1.08 g) was heated at refluxin a mixture of EtOH (100 mL) and water (10 mL) for 4 hours. Aftercooling to room temperature the precipitate was collected by filtration,washed with water (2×50 mL), and dried under vacuum to give4,5-dimethoxy-1-indanone oxime (0.80 g, 72%).

To a degassed solution of the crude oxime (from step 1, 0.80 g, 3.80mmol) in EtOH/AcOH (9/1, 50 mL) was added Pd—C (10%, 100 mg). Themixture was hydrogenated under an atmospheric hydrogen pressureovernight, and then filtrated through Celite™. The filtrate wasconcentrated and the residual material was azeotroped twice with tolueneto yield 4,5-dimethoxyindan-1-amine as the acetate salt, employed in thenext step without further purification.

A mixture of the 4,5-dimethoxyindan-1-amine acetic acid (0.93 g, 3.67mmol) and the sultone (0.49 g, 3.67 mmol) in mixture ofacetonitrile/toluene (15 mL/5 mL) was heated at reflux for 5 hours.After cooling to room temperature the solid formed was collected byfiltration, and then suspended in EtOH (40 mL). The suspension washeated at reflux for 1 hour. The mixture was cooled to room temperature;the solid material was collected by filtration, washed with acetone(2×20 mL) and dried in vacuo to afford the title compound (0.75 g, 65%);¹H NMR (CD₃OD, 500 MHz) δ ppm 7.09 (d, 1H, J=8.5 Hz), 6.87 (d, 1H, J=8.0Hz), 4.18 (t, 1H, J=6.5 Hz), 3.32 (s, 3H), 3.31 (s, 3H), 3.04 (m, 1H),2.88 (m, 1H), 2.78 (m, 3H), 2.34 (m, 1H), 2H), 2.02 (m, 2H), 1.91 (m,1H); ES-MS 314 (M−1).

Preparation of 2-(aminomethyl)-3,3-dimethylbutane-1-sulfonic acid:(Compound N78)

To a solution of 4-tert-butyl-1,2-oxathiolane 2,2-dioxide (468 mg, 26.3mmol) in dimethylformamide (4 mL) was added sodium azide (187 mg). Thesolution was heated at 130° C. for 36 hours. The reaction mixture wascooled to room temperature and the solvent was evaporated under vacuum.The residue was triturated with ethanol; and the solid material wascollected by filtration and washed with acetone (2×20 mL) to give thecorresponding azido derivative (420 mg, 72%).

To a degassed solution of the above obtained azido derivative (400 mg)in EtOH (75 mL) was added Pd—C (10%, 80 mg). The mixture washydrogenated under an atmospheric hydrogen pressure for 24 hours, andthen filtrated through Celite™ to give sodium salt of the title compoundquantitatively. The salt was dissolved in MeOH (20 mL); and themethanolic solution was treated with Amberlite® 1R-120(plus) ionexchange resin (4 g, pre-washed with methanol and water). The mixturewas stirred at room temperature for 30 minutes, and the insolublematerials was removed by filtration. Evaporation of the solvent gave thetitle compound as a white solid (400 mg, 98%); ¹H NMR (CD₃OD, 500 MHz) δppm 3.08 (m, H), 2.95 (m, 1H), 2.74 (m, 1H), 2.63 (m, 1H), 1.76 (br t,1H, J=9.5 Hz), 0.95 (s, 9H); ES-MS 194 (M−1).

Preparation of (1E,3S)-3-amino-5-methylhex-1-ene-1-sulfonic acid(Compound N79)

In a dry 500-mL three necked round-bottomed flask equipped with magneticstirring bar, additional funnel and a thermometer, was addedN-Boc.Leu.OMe (6.0 g, 23.4 mmol) under nitrogen. Dry toluene was addedand the solution was cooled to −78° C. DIBAL-H 1M solution in toluene(36 mL, 36 mmol) was added dropwise to the reaction mixture over 1 hour.The rate of addition was adjusted so that the internal temperature wasmaintained below −65° C. The reaction mixture was then stirred for anadditional 2 h at −78° C. under nitrogen. The reaction was quenched byadding slowly 5 mL of cold MeOH (−78° C.). Again the internaltemperature was kept below −65° C. The reaction mixture was thencarefully poured into a cold solution of HCl 1N (100 mL) with stirringover 15 minutes. The aqueous mixture was then extracted with EtOAc(3×150 mL). The organic layers were combined, washed with brine (3×100mL), and dried over MgSO₄. The solvent was removed by evaporation underreduced pressure to give of the corresponding aldehyde quantitatively,which was used in the next step without purification.

Ethyl (diethoxyphosphoryl)methanesulfonate (5.05, 19.4 mmol) was addedto a suspension of sodium hydride (400 mg, 16.7 mmol) in dry THF (100mL) at 0° C. The reaction mixture was stirred at 0° C. for 15 minutes,followed by addition of the aldehyde prepared in step 1 (4.8 g, 22mmol). The reaction mixture was stirred at 0° C. for 2 hours, and thenat room temperature for 30 minutes. The reaction mixture was poured intowater, extracted with EtOAc (3×100 mL). The extracts were combined,washed with brine, dried over MgSO₄, and concentrated to dryness. Theoily residue was purified by Biotage chromatography using EtOAc/hexane(1:1), providing the corresponding unsaturated sulfonic acid ethyl esteras a colorless oil which crystallized on a standing in vacuo. (5.43 g,77% yield).

The ester product from step 2 (1.0 g, 4.6 mmol) was dissolved in formicacid (10 mL) and water (0.5 mL). The mixture was heated at 50° C. for 24hours, cooled to room temperature, and concentrated under reducedpressure. The resultant residue was dried in vacuo. The gum-likematerial was triturated in ethanol and turned into solid. The solid wascollected by filtration, washed with ethanol to give the title compound(0.495 g, 82%); ¹H NMR (CD₃OD, 500 MHz) δ ppm 6.66 (d, 1H, J=15 Hz),6.32 (dd, 1H, 8.3 & 7.3 Hz), 3.88-3.93 (m, 1H), 1.52-1.65 (m, 3H), 0.97(dd, 6H, J=6 and 7.0 Hz); ES-MS 192 (M−1).

Preparation of 3-[(6-methylindan-1-yl)amino)propane-1-sulfonic acid(Compound N81)

A mixture of 6-methyl-1-indanone (0.97 g, 6.64 mmol), hydroxylaminehydrochloride (1.11 g) and sodium acetate (1.31 g) was heated at refluxin ethanol/water (50 mL/1.5 mL) for 5 hours. After cooling to roomtemperature the resulting precipitate was filtered and washed with water(2×50 mL) and dried in vacuo to give 6-methyl-1-indanone oxime (0.98,91%) oxime which was used in the next step without purification.

To a degassed a solution of the crude oxime (from step 1, 1 g, 6.2 mmol)in EtOH/AcOH (9/1, 50 mL) was added Pd—C (10%, 200 mg). The mixture washydrogenated under an atmospheric hydrogen pressure overnight, and thenfiltrated through Celite™. The filtrate was concentrated and the residuewas azeotroped twice with toluene to yield 6-methylindan-1-amine as theacetate salt (1.24 g, quantitative), which was employed in the next stepwithout purification.

The a mixture of the 6-methylindan-1-amine acetic acid (1.2 g, 6.15mmol) and 1,3-propane sultone (0.75 g, 5.5 mmol) in acetonitrile/toluenemixture (15 mL/5 mL) was refluxed for 5 hours. After cooling to roomtemperature the white solid obtained was collected by filtration, andwas suspended in EtOH (40 mL). The suspension was stirred at reflux for1 hour. The mixture was cooled to room temperature; the solid materialwas collected by filtration, washed with acetone (2×20 mL) and dried invacuo to afford the title compound (0.85 g, 53% yield); ¹H NMR (CD₃OD,500 MHz) δ ppm 7.20 (s, 1H), 7.08 (d, 1H, J=7.5 Hz) 7.00 (d, 1H, J=5.3Hz), 4.20 (t, 1H, J=7.0 Hz), 2.87 (m, 1H), 2.78 (m, 3H), 2.36 (m, 1H),2.31 (s, 3H), 2H), 2.02 (m, 2H), 1.84 (m, 1H). ES-MS 268 (M−1).

Preparation of (3S)-3-amino-5-methylhexane-1-sulfonic acid (CompoundN82)

To a degassed a solution of3-(t-butyloxycarbamido)-5-methylhex-1-ene-1-sulfonic acid ethyl ester(1.0 g, 2 mmol) in EtOH/AcOH (9/1, 50 mL) was added Pd—C (10%, 100 mg).The mixture was hydrogenated under an atmospheric hydrogen pressure for1.5 hours, and then it was flushed with nitrogen and filtrated throughCelite™. The celite pad was washed with ethanol several times. Thefiltrate and the washings were combined; and the solvent was removed byevaporation, providing 3-(t-butyloxycarbamido)-5-methylhexane-1-sulfonicacid ethyl ester (0.99 g, 98%) which was used in the next step withoutpurification.

3-(t-Butyloxycarbamido)-5-methylhexane-1-sulfonic acid ethyl ester (1.0g, 3.0 mmol) was treated in formic acid (10 mL) and water (0.5 mL) at50° C. for 24 hours. The reaction mixture was concentrated and dried invacuo. The gum-like material was triturated in ethanol. The solid wascollected by filtration, washed with ethanol to give the title compound(0.430 g, 72%); ¹H NMR (CD₃OD, 500 MHz) δ ppm 3.39-3.36 (m, 1H),3.45-3.41 (m, 1H), 2.95-2.91 (m, 2H) 2.04-1.98 (m, 1H), 1.74-1.70 (m,1H), 1.56-1.41 (m, 2H), 0.98 (dd, 6H, J=3.4 and 7.2 Hz); ES-MS 194(M−1).

Preparation of Compounds N38, N40, and N41

These three compounds were prepared from a common starting material,namely 2-(t-butyl)-1,3-propane sultone, which was prepared as thefollowing: To a stirred solution of lithium bis(trimethylsilyl)amide1.0M solution in THF (42 mL, 42 mmol) at −78° C. was added dropwise asolution of ethyl methanesulfonate (4.30 mL, 10 mmol) in dry THF (5 mL).The mixture was stirred at −78° C. for 30 minutes then bromopinacolone(5 g, 27.9 mmol) in dry THF (10 mL) was added dropwise. The mixture wasthen stirred at −78° C. for 2 hours, and at −50° C. for 2 hours, andfinally quenched with a saturated solution of ammonium chloride (100mL). The mixture was extracted with EtOAc (3×100 mL). The organic layerswere combined, washed with water (2×50 mL), brine (100 mL), dried overMgSO₄, and concentrated under reduced pressure. The residual materialwas purified by Biotage using Hexane:EtOAc (9:1) to give2-(t-butyl)-1,3-prop-1-ene sultone (3.78 g, 75%). To a degassed asolution of 2-(t-butyl)-1,3-prop-1-ene sultone (1 g, 5.67 mmol) in MeOH(50 mL) was added 10% Pd—C (10%, 150 mg). The mixture was hydrogenatedunder 40 Psi hydrogen pressure for 48 h. The mixture was filtratedthrough Celite™ and the filtrate was concentrated to give2-(t-butyl)-1,3-propane sultone (0.90 g, 89%).

2[(t-Butylamino)methyl]-3,3-dimethylbutane-1-sulfonic acid (CompoundN38)

To a solution of 2-(t-butyl)-1,3-propane sultone (83 mg, 0.466 mmol) indimethylformamide (2 mL) was added t-butylamine (2 mL). The solution wasstirred at 130° C. for 48 hours. The reaction mixture was cooled to roomtemperature and the solvent was removed under reduced pressure. Theresidue was triturated with ethanol; and the solid material wascollected by filtration, washed successively with ethanol (2×20 mL) andwith acetone (2×20 mL), and dried under high vacuum to afford the titlecompound (56 mg, 62%); ¹H NMR (CD₃OD, 500 MHz) δ ppm 3.29 (m, 2H), 2.98(m, 1H), 2.82 (m, 1H), 1.98 (br t, 1H, J=9.5 Hz), 1.36 (s, 9H), 1.05 (s,9H); ES-MS 250 (M−1).

2-[(1-Adamantylamino)methyl]-3,3-dimethylbutane-1-sulfonic acid(Compound N40)

To a solution of 2-(t-butyl)-1,3-propane sultone (240 mg, 1.34 mmol) indimethylformamide (10 mL) was added 1-adamanthanamine (204 mg, 1.34mmol). The solution was heated at 130° C. for 24 hours. The reactionmixture was cooled to room temperature and was triturated with ethanol.The solid was collected by filtration, washed successively with ethanol(2×20 mL) and acetone (2×20 mL), and purified by LC-MS to provide thetitle compound (25 mg, 5%); ¹H NMR (CD₃OD, 500 MHz) δ ppm 3.00 (m, 2H),2.98 (m, 1H), 2.81 (m, 1H), 2.21 (br s, 3H), 1.94 (br t, 1H, J=9.5 Hz),1.92 (m, 6H), 1.75 (m, 6H), 1.02 (s, 9H); ES-MS 328 (M−1).

3,3-Dimethyl-2-[(methylamino)methyl]butane-1-sulfonic acid: (CompoundN41)

A mixture of 2-(t-butyl)-1,3-propane sultone (70 mg, 0.39 mmol) andexcess of methylamine 1M in THF (5 mL) was heated in a metal cylinder at130-° C. for 72 hours. The reaction mixture was cooled to roomtemperature and the solid material was collected by filtration andwashed with acetone (2×20 mL) to give the title compound (50 mg, 62%);¹H NMR (CD₃OD, 500 MHz) δ ppm ¹H NMR (CD₃OD, 500 MHz) δ ppm 3.13 (m,1H), 2.79 (m, 1H), 2.70 (m, 1H), 2.53 (m, 1H), 2.35 (s, 3H), 1.86 (m,1H), 0.98 (s, 9H); ES-MS 208 (M−1).

Preparation of (3S)-3-amino-5-methylhexane-1-sulfonic acid: (CompoundN89)

To a 1.0 M stirred solution of lithium bis(trimethylsilyl)amide in THF(9 mL, 9 mmol) at −78° C. was added dropwise a solution of ethyl(3S)-3-[(tert-butoxycarbonyl)amino]-5-methylhexane-1-sulfonate (1.96 g,6.0 mmol) in dry THF (50 mL). The mixture was stirred at −78° C. for 30minutes followed by dropwise addition of methyl iodide (0.47 mL, 7.6mmol) in dry THF (10 mL). The mixture was stirred at −78° C. for 4hours, and then quenched with a saturated solution of ammonium chloride(10 mL). The mixture was extracted with EtOAc (3×50 mL). The combinedorganic phase was washed with water (2×50 mL), brine (100 mL), driedover MgSO₄ and concentrated in vacuo to give colorless oil. Theresultant oil was purified by Biotage chromatography using EtOAc/Hexane1/5, to giving the corresponding α-methylated intermediate (0.85 g,42%). The methylated intermediate (0.85 g, 2.5 mmol) was treated withformic acid (5 mL) and water (0.25 mL) at 50° C. for 24 hours. Thereaction mixture was concentrated and dried in vacuo. The gum-likematerial thus obtained was triturated in ethanol. The solid material wascollected by filtration and washed with ethanol, providing the titlecompound (0.273 g, 54%); ¹H NMR (CD₃OD, 500 MHz) δ ppm 3.47-3.45 (1 m,1H), 3.45-3.41 (m, 1H), 2.99-2.91 (m, 1H), 2.24-2.18 (m, 1H), 2.09-2.03(m, 1H), 1.86-1.80 (m, 2H), 1.78-1.69 (m, 2H), 1.32 (d, 3H, J=4.3), 0.98(dd, 6H, J=3.9 and 4.2 Hz); ES-MS 208 (M−1).

Preparation of 2-(1-adamantyl)-3-(tert-butylamino)propane-1-sulfonicacid (Compound N90) and2-(1-adamantyl)-3-(methylamino)propane-1-sulfonic acid (Compound N91)

A solution of ethylmethane sulfonate (1.21 mL, 11.7 mmol) in THF (10 mL)was cooled to −78° C. To the cold solution was added dropwise LiHMDS 1Msolution in THF (12.5 mL, 12.5 mmol). The mixture was stirred for 30minutes, and then transferred via cannula to a second flask containing asolution of adamanthyl bromomethyl ketone (2.00 g, 7.80 mmol) in THF (40mL) at −78° C. under N₂ atmosphere. The resultant mixture was stirred at−78° C. for 1.5 hours and then warmed slowly to −45° C., and stirred for2.5 hours at this temperature. The reaction was quenched by adding asaturated solution of NH₄Cl (100 mL). The mixture was extracted withEtOAc (3×100 mL) and washed with brine, dried over MgSO₄ andconcentrated under reduced pressure to obtain a yellow solid (1.93 g,97%). The solid (0.66 g, 2.59 mmol) was hydrogenated in the presence ofpalladium on activated charcoal (10%, 0.198 g) in MeOH (60 mL), in aPyrex™ high pressure bottle and hydrogenated at 40 psi for 48 h. Thereaction mixture was filtered through Celite™ and concentrated to give2-(1-admantyl)-1,3-propane sultone as a white solid (0.65 g, 98%).

2-(1-Adamantyl)-3-(tert-butylamino)propane-1-sulfonic acid (CompoundN90)

The above-derived 2-(1-adamantyl)-1,3-propane sultone (0.12 g, 0.466mmol) was dissolved in DMF, followed by addition of t-butylamine (0.074mL, 0.699 mmol). The mixture was heated at 130° C. Additionalt-butylamine (0.07 mL, 0.663 mmol) was added to the reaction mixture.The next day, reaction mixture was cooled to room temperature and moret-butylamine (1.0 mL) was added and the resulting mixture was heatedagain 130° C. After being stirred for 8 hours, some more t-butylamine (3mL) was added and the reaction mixture was heated for 3 more days at130° C. The reaction mixture was concentrated by evaporation, causing awhite solid to precipitate. The mixture was diluted with MeOH (1 mL);and the white solid was collected by filtration, washed with Et₂O, anddried in vacuo, providing the title compound (0.062 g, 51%); ¹H NMR(CD₃OD, 500 MHz) δ ppm 3.36-3.32 (m, 2H), 2.93 (dd, 1H, J=9.8 Hz,J=12.7), 2.75 (dd, 1H, J=9.3 Hz, J=14.6 Hz), 2.02 (large s, 3H),1.78-1.50 (m, 13H), 1.37 (s, 9H). ¹³C NMR (CD₃OD, 125 MHz) δ ppm 56.70,51.71, 48.43, 43.53, 43.31, 38.87, 36.58, 35.52, 28.75, 24.67. ES-MS:328 (M−1).

2-(1-Adamantyl)-3-(methylamino)propane-1-sulfonic acid (Compound N91)

2-(1-Adamantyl)-1,3-propane sultone (0.104 g, 0.405 mmol) was introducedinto a Pyrex™ high pressure bottle and a solution of 2 M methylamine inTHF (8 mL) was added. The bottle was closed and heated at 110° C.overnight, and then cooled to room temperature. The white precipitatewas collected by filtration, washed with Et₂O, and dried in vacuo,giving the title compound (0.0681 g, 59%); ¹H NMR (CD₃OD, 500 MHz) δ ppm4.39-4.36 (m, 1H), 3.27-3.24 (m, 1H), 3.0 (dd, 1H, J=9.3 Hz, J=12.7 Hz),2.78 (dd, 1H, J=9.8 Hz, J=14.2 Hz), 2.71 (s, 3H), 2.02 (large s, 3H),1.80-1.57 (m, 13H); ES-MS: 286 (M−1).

Example 2 Binding and Antifibrillogenic Assays

A representative number of compounds according to the invention weresynthesized and screened by mass spectrometry (“MS”) assays, except forselected compounds which were purchased from a commercial source. The MSassay gives data on the ability of selected compounds to bind toproteins, in this example, to Aβ40.

Samples were prepared as aqueous solutions (adding 20% ethanol ifnecessary to solubilize in water). For those compounds selected fromTable 2A, the binding experiment was done with 200 μM of a test compoundand 20 μM of solubilized Aβ40, or 400 μM of a test compound and 40 μM ofsolubilized Aβ40. For those compounds selected from Table 3A and Table3B, the binding experiment was done with 150 μM of a test compound and30 μM of solubilized Aβ40. The pH value of each sample was adjusted to7.4 (±0.2) by addition of 0.1% aqueous sodium hydroxide. The solutionswere then analyzed by electrospray ionization mass spectrometry using aWaters ZQ 4000™ mass spectrometer. Samples were introduced by directinfusion at a flow-rate of 25 μL/min within 2 hours after samplepreparation. The source temperature was kept at 70° C. and the conevoltage was 20 V for all the analysis. Data were processed usingMasslynx 3.5™ software. The MS assay gives data on the ability ofcompounds to bind to soluble Aβ, whereas the ThT, EM and CD assays givedata on inhibition of fibrillogenesis. The results from the assay forbinding to Aβ for compounds selected from Table 2A are summarized inTable 4 and the results from the assay for binding to Aβ for compoundsselected from Tables 3A and 3B are summarized in Table 5.

TABLE 4 Relative Binding Affinities of Selected Compounds Shown in Table2A Aβ binding (%) Compound ID 90-100% @ AY; BB; BV; BW; BX; BY; BZ; CE;CG; CH; CI; CJ; 400 μM; CK; CV; CY; DC; DD; DK; DO; DU; DV; DX; DY; DZ;60-100% @ EB; ED; EE; EG; EH; EK; ES; ET; EY; EZ; FA; FS; FX; 200 μM FY;NJ; 70-89% @ AG; AK; AL; AW; AX; AZ; BA; CC; CD; DH; DM; DN; 400 μM; DW;EA; EJ; EL; ER; FP; FR; FU; GD; GJ; GL; I; NG; 30-69% @ 200 μM 45-59% @AC; AD; AE; AF; AH; AM; B; BC; C; D; DG; DL; DP; 400 μM; DQ; DR; DS; DT;E; EF; EN; EO; EP; EQ; F; FQ; FZ; 20-44%@ G; GB; GH; GI; GK; GS; GU; H;HC; HG; J; L; NH; NI; 200 μM NK; 20-39% @ DE; DF; DI; DJ; EI; FH; FO;FT; FV; FW; GA; GC; GM; 400 μM and GN; GO; GP; GQ; GR; GT; GZ; HA; HB;HD; HE; HF; 200 μM HJ; HK; K; M; N; P; Q

TABLE 5 Relative Binding Affinities of Selected Compounds Shown inTables 3A and 3B Aβ binding (%) Compound ID  ≧70% OE; OM; OV; OW; OY;PD; PI; PK; PT; PW; QG; QT; QU; QW; SX; SY; SZ; N1; N4; N10; N11; N12;N16; N17; N24; N25; N30; N33; N36; N42; N44; N52; N53; N54; N56; N57;N58; N61; N64; N65; N66; N73; N74; N78; N79 40-69% NN; NM; NO; NP; NR;NU; NV; NZ; OA; OB; OC; OD; OI; OL; ON; OO; OP; OQ; OR; OU; OX; OZ; PA;PE; PF; PG; PH; PJ; PL; PQ; PR; PS; PV; PY; QB; QC; QD; QE; QF; QH; QI;QJ; QL; QM; QN; QP; QQ; QR; QS; QV; QX; N2; N5; N7; N8; N9; N13; N14;N18; N19; N20; N23; N26; N31; N32; N34; N35; N37; N38; N39; N40; N41;N43; N47; N48; N49; N50; N51; N55; N59; N60; N62; N63; N69; N70; N71;N72; N77; N82; N84; N85; N87; 15-39% NQ; NS; NT; NW; NX; NY; OF; OG; OH;OJ; OK; OS; OT; PB; PM; PN; PO; PP; PU; PX; PZ; QA; QK; QO; N3; N6; N15;N21; N22; N27;

Example 3 Effects of Short Term Treatment in Adult Transgenic CRND8 MiceOverexpressing βAPP

Transgenic mice, TgCRND8, expressing the human amyloid precursor protein(hAPP) develop a pathology resembling Alzheimer's disease. Inparticular, high levels of Aβ40 and Aβ42 have been documented in theplasma and the brain of these animals at 8-9 weeks of age, followed byearly accumulation of amyloid plaques similar to the senile plaquesobserved in AD patients. These animals also display progressivecognitive deficits that parallel the appearance of degenerative changes.See, e.g., (Chishti, et al., J. Biol. Chem. 276, 21562-70 (2001).

The short term therapeutic effect of 19 compounds of the invention wasstudied. These compounds were administered over a 14 or 28 day period atthe end of which the levels of Aβ peptides in the plasma and brain ofTgCRND8 animals were determined.

Methods

Male and female transgenic mice from the 3^(rd) and 4^(th) B6C3F1generations were used in this example and given daily subcutaneous ororal administrations of one of a series of compounds for 14 or 28 days.The following abbreviations are used to designate these animals from the3^(rd) and 4th generation backcross in the present protocol:TgCRND8-2.B6C3F1(N₃); TgCRND8-2.B6C3F1 (N₄).

Baseline animals (Group 1) consisted of naive TgCRND8-2. B6C3F1(N₃) at11±1 weeks of age. These mice were used to determine the Aβ levels inthe plasma and brain of naive transgenic animals at the initiation oftreatment.

Starting at 11 weeks of age (±1 week) animals received dailyadministration of their respective treatment for a period of 14 or 28days (groups 2-21), at a dose of 250 mg/kg at 10 ml/kg or of vehicleonly (water; group 2) or 1% methyl cellulose only (group 21). The routeof administration was subcutaneous for water-soluble compounds and oralfor compounds solubilized in methylcellulose 1% (MC 1%). At the end ofthe treatment periods, plasma and perfused brains were collected forquantification of Aβ levels.

The mice used in this study were derived from a breeding colony atInstitut Armand Frappier, and were well-acclimated to the animalfacility environment prior to initiation of the study. Animals wereassigned, according to age and gender, into the following experimentalgroups:

TABLE 6 Groups of Mice Daily Dose Duration of Treatment Group No.Treatment (mg/kg) (days) 1 Baseline NA NA 2 Water NA 14 & 28 4 BY 250 14& 28 6 CV 250 14 & 28 12 CY NA 14 & 28 15 BW 250 14 & 28 16 BZ 250 14 &28 18 BX 250 14 & 28 20 DC 250 14 & 28 21 Methylcellulose 1% 100 14 & 2822 DD 250 14 &28  23 DH 250 14 & 28 24 DM 250 14 &28  25 DX 250 14 &28 26 DY 250 14 &28  27 DZ 250 14 &28  28 ED 250 14 &28  29 EG 250 14 &28 

Animal Health Monitoring

All animals were examined daily for signs of ill health when handled inthe morning for their daily treatment and twice a day for mortalitychecks (once daily during weekends and holidays). Detailed examinationswere performed on the treatment initiation, weekly during the study, andonce before terminal procedures. More frequent observations wereundertaken when considered appropriate. Death and all individualclinical signs were individually recorded. Individual body weights wererecorded at randomization, once weekly during the study, and once beforeterminal procedures.

Sample Collection

At 11±1 weeks of age for the Baseline group, and at the end of thetreatment period (14 or 28 days) for Groups 2 to 21, at 24 hours afterthe last compound administration animals were sacrificed and samplescollected. An approximate blood volume of 500 μl was collected from theorbital sinus and kept on ice until centrifugation at 4° C. at a minimumspeed of 3,000 rpm for 10 minutes. Plasma samples were immediatelyfrozen and stored at −80° C. pending analysis. The brains were removed,frozen, and stored at −80° C. awaiting analysis.

Measurements of Aβ Levels

Brains were weighted frozen and homogenized with 4 volumes of ice cold50 mM Tris-Cl pH 8.0 buffer with protease inhibitor cocktail (4 mL ofbuffer for 1 g of wet brain). Samples were spun at 15000 g for 20minutes and the supernatants were transferred to fresh tubes. Onehundred fifty (150) μl from each supernatant were mixed with 250 μl of8M guanidine-HCL/50 mM Tris-HCL pH 8.0 (ratio of 0.6 vol supernatant: 1vol 8M guanidium/Tris-HCL 50 mM pH8.0) and 400 μL 5 M guanidium/Tris-HCL50 mM pH8.0 were added. The tubes were vortexed for 30 seconds andfrozen at −80° C. In parallel, pellets were treated with 7 volumes of 5M guanidine-HCL/50 mM Tris-HCL pH 8.0 (7 mL of guanidine for 1 g of wetbrain), vortexed for 30 seconds and frozen at −80° C. Samples werethawed at room temperature, sonicated at 80° C. for 15 minutes andfrozen again. This cycle was repeated 3 times to ensure homogeneity andsamples were returned to −80° C. pending analysis.

Aβ levels were evaluated in plasma and brain samples by ELISA usingHuman Aβ40 and Aβ42 Fluorometric ELISA kits from Biosource (Cat. No.89-344 and 89-348) according to manufacturer's recommended procedures.In short, samples were thawed at room temperature, sonicated for 5minutes at 80° C. (sonication for brain homogenates; no sonication forplasma samples) and kept on ice. Aβ peptides were captured using 100 μlof the diluted samples to the plate and incubated without shaking at 4°C. overnight. The samples were aspirated and the wells were rinsed 4times with wash buffer obtained from the Biosource ELISA kit. Theanti-Aβ40 or anti-Aβ42 rabbit polyclonal antiserum (specific for theAβ40 or Aβ42 peptide) was added (100 μl) and the plate was incubated atroom temperature for 2 hours with shaking. The wells were aspirated andwashed 4 times before adding 100 μl of the alkaline phosphatase labeledanti-rabbit antibody and incubating at room temperature for 2 hours withshaking. The plates were then rinsed 5 times and the fluorescentsubstrate (100 μl) was added to the plate. The plate was incubated for35 minutes at room temperature and the plate was read using a titerplate reader at an excitation wavelength of 460 nm and emission at 560nm.

Compounds were scored based on their ability to modulate levels of Aβpeptides in the plasma and the cerebral soluble/insoluble levels in thebrain. Levels of Aβ observed in the plasma and brain of treated animalswere normalized using values from vehicle-treated (water) ormethylcellulose-treated control groups and ranked according to thestrength of the pharmacological effect. Results are shown in Tables 3 to11. Increases in the levels of Aβ peptides are indicated using “+”symbols, while decreases in the levels of Aβ peptides are indicatedusing “−” symbols. The strongest effects are recorded as “+++” or “−−−”while the weakest are shown as “+” or “−”.

Specifically, increases in the levels of Aβ (relative to untreatedcontrol) of 20 to 39% are scored as “+”; increases of 40 to 69% arescored as “++”; and increases of 70% or higher are scored as “+++”.Decreases in the levels of Aβ of 5 to 19% are scored as “−”; decreasesof 20 to 38% are scored as “−−”; and decreases of 39% or more are scoredas “−−−”.

The data are presented in Tables 6-11. Treatment with these compoundsafter 14 and/or 28 days resulted in a significant change in the cerebrallevels of Aβ40 and/or Aβ42 (Tables 8-11). Furthermore, treatment withthese compounds, for instance, Compound BX(3-(t-butyl)amino-1-propanesulfonic acid), resulted after 14 and 28 days(Tables 6-7) in a significant increase in the levels of Aβ peptides inthe plasma. This suggests that some of these compounds may act by aperipheral sink effect, sequestering Aβ peptides in the plasma andthereby facilitating their clearance from the CNS as previouslysuggested for treatment by passive immunization using anti-Aflmonoclonal antibody m266 (DeMattos et al., Science 295(5563):2264-7).

The tables show levels of Aβ peptides in the plasma and brain of TgCRND8mice treated for 14 and 28 days with compounds of the invention.

Tables 6A and 6C show the data from Day 14 and Day 28 for the PlasmaVehicle group, respectively. Tables 6B and 7 show the data for thePlasma Methylcellulose group on Days 14 and 28, respectively. Tables 8and 10 show the data on Days 14 and 28 for the Brain homogenate vehiclegroup, respectively. Tables 9 and 11 show the data for brain homogenatefor the Methylcellulose group on Days 14 and 28, respectively.

TABLE 6A Plasma Vehicle Group, Day 14 Treatment Aβ40 Change Aβ42 ChangeBY + + CV + ++ DC ++ ++ BX +++ ++

TABLE 6B Plasma Methylcellulose Group, Day 14 Treatment Aβ40 Change Aβ42Change BZ + BW CY

TABLE 6C Plasma Vehicle Group, Day 28 Treatment Aβ40 Change Aβ42 ChangeBY CV ++ ++ DC ++ BX +++

TABLE 7 Plasma Methylcellulose Group, Day 28 Treatment Aβ40 Change Aβ42Change BZ ++ ++ BW + CY +

TABLE 8 Brain Homogenate Vehicle Group, Day 14 Aβ40 Change Aβ42 ChangeTreatment Soluble Insoluble Soluble Insoluble BY +++ +++ +++ CV** − DC−− + −− BX − −−− −− −−− DD − DX −− − DY −− −− DZ −− EG − −− −− −− DH −−−−− −−− DM + − − ED − + **The effect of this compound in the brain hasonly been tested on its ability to modulate the total levels of Aβ40 andAβ42 peptides rather than measuring soluble and insoluble levelsindependently.

TABLE 9 Brain Homogenate Methylcellulose Group, Day 14 Aβ40 Change Aβ42Change Treatment Soluble Insoluble Soluble Insoluble BZ −−− −− BW −−− −−−−− CY − +++ ++

TABLE 10 Brain Homogenate Vehicle Group, Day 28 Aβ40 Change Aβ42 ChangeTreatment Soluble Insoluble Soluble Insoluble BY + +++ +++ CV** ++ +++DC − + ++ +++ BX −−− −−− −− − DD − DX −− − −− DY −−− − −− DZ − −− −− −−EG −− −− DH − − DM − −− −− −− ED − −− − − **The effect of this compoundin the brain has only been tested on its ability to modulate the totallevels of Aβ40 and 42 peptides rather than measuring soluble andinsoluble levels independently.

TABLE 11 Brain Homogenate Methylcellulose Group, Day 28 Aβ40 Change Aβ42Change Treatment Soluble Insoluble Soluble Insoluble BZ −− −− −− BW − −− −− CY ++ +++ +++ +

Example 4 Effects of Long Term Treatment in Adult Transgenic CRND8 MiceOverexpressing βAPP

Transgenic mice, TgCRND8, as those used in the short term treatment,overexpress a human APP gene with the Swedish and Indiana mutationsleading to the production of high levels of the amyloid peptides and tothe development of an early-onset, aggressive development of brainamyloidosis. The high levels of Aβ peptides and the relativeoverabundance of Aβ₄₂ compared to Aβ₄₀ are believed to be associatedwith the severe and early degenerative pathology observed. The patternof amyloid deposition, presence of dystrophic neuritis, and cognitivedeficit has been well documented in this transgenic mouse line. Thelevels of Aβ peptides in the brain of these mice increase dramaticallyas the animals age. While the total amyloid peptide levels increase from˜1.6×10⁵ pg/g of brain to ˜3.8×10⁶ between 9 and 17 weeks of age.

While the early deposition of amyloid in this model allows the rapidtesting of compounds in a relatively short time frame, the aggressivityof this model and the high levels of Aβ peptides renders therapeuticassessment in the longer term a more difficult task.

The long-term therapeutic effects of compounds of the present inventionon cerebral amyloid deposition and β-amyloid (Aβ) levels in the plasmaand in the brains of transgenic mice, TgCRND8, expressing the humanamyloid precursor protein (hAPP) was studied. These compounds wereadministered over an 8 or 16 week period at the end of which the levelsof Aβ peptides in the plasma and brain of TgCRND8 animals weredetermined. The goal of this study was to evaluate the efficacy of thecompounds at modulating the progression of the amyloidogenic process inthe brain and in the plasma of a transgenic mouse model of Alzheimer'sdisease (AD)

Methods

The mice used in the study consisted of animals bearing one copy of thehAPP gene (+/−) from the 2^(nd) and 3^(rd) generation progenies (N₂ andN₃) derived from backcrosses from TgCRND8.FVB(N₂)AJ(N₃) with B6AF1/Jhybrid animals.

-   -   N₁=TgCRND8.FVB(N₂)AJ(N₃)×B6AF1/J    -   N₂=TgCRND8.FVB(N₂)AJ(N₃).B6AF1/J(N₁)×B6AF1/J    -   N₃=TgCRND8.FVB(N₂)AJ(N₃).B6AF1/J(N₂)×B6AF1/J

The following abbreviations are used to designate these animals in thepresent study: TgCRND8.B6AF1/J(N₂); TgCRND8.B6AF1/J(N₃). Male and femaletransgenic mice were given daily subcutaneous (compound BX) or oral(compounds BW and BZ) administrations of the appropriate compounds for 8or 16 weeks.

Baseline animals consisted of 9±1 week old naive TgCRND8.B6AF1/J animalsfrom the 2^(nd) and 3^(rd) generations. These mice were used todetermine the extent of cerebral amyloid deposits and Aβ levels in theplasma and brain of naive transgenic animals at the initiation oftreatment.

Starting at 9 weeks of age (±1 week) animals received dailyadministration of their respective treatment for a period of 8 or 16weeks, at a dose of 30 or 100 mg/kg at 10 ml/kg. The route ofadministration was subcutaneous for water-soluble compounds (CompoundBX) and oral for compounds solubilized in methylcellulose 1% (MC 1%)(Compounds BW and BZ). At the end of the treatment periods, plasma andperfused brains were collected for quantification of Aβ levels.

Animal health was monitored, samples were collected and Aβ levels weremeasured as described above in the short term treatment study. Compoundswere scored based on their ability to modulate levels of Aβ peptides inthe plasma and the cerebral soluble/insoluble levels in the brain.Levels of Aβ observed in the plasma and brain of treated animals werecompared to that of vehicle-treated (water) or methylcellulose-treatedcontrol groups and ranked according to the strength of thepharmacological effect. Results are shown in Table 12. Increases in thelevels of Aβ peptides are indicated using “+” symbols, while decreasesin the levels of Aβ peptides are indicated using “−” symbols. Thestrongest effects are recorded as “+++” or “−−−” while the weakest areshown as “+” or “−”.

Specifically, increases in the levels of Aβ (relative to vehicle treatedcontrol) of 5 to 14% are scored as “+”; increases of 15 to 29% arescored as “++”; and increases of 30% or higher are scored as “+++”.Decreases in the levels of Aβ of 5 to 14% are scored as “−”; decreasesof 15 to 29% are scored as “−−”; and decreases of 30% or more are scoredas “−−−”. Additionally, changes of 4% or less in either direction arescored as “0”.

Table 12 shows levels of Aβ peptides in the plasma and brain of TgCRND8mice treated for 8 and 16 weeks with compounds of the invention.Treatment with these compounds after 8 and/or 16 weeks in many casesresulted in a change in the levels of Aβ₄₀ and/or Aβ₄₂ in the plasmaand/or brain. For example, administration of compound BX generallyresulted in a dramatic decrease in the amount of Aβ in the brain at both8 and 16 weeks. Compound BW also resulted in a dramatic decrease inbrain and plasma levels of Aβ at 8 weeks and plasma levels at 16 weeks.

For the ThioS studies, the plaques in the brains of the mice werequantified as follows. Mice were transcardially perfused with salinesolution. Brains were dissected out and separated in 2 hemispheres. Theleft hemisphere was immersed in freshly-prepared 4% paraformaldehyde for3 hrs at 4° C., then transferred into 30% sucrose at 4° C. Whencryoprotection was achieved (24-48 hours), brains were frozen inisopentane at −45° C. and subsequently stored at −80° C. untilsectioning. Coronal 40 μm-thick sections were performed, and stainedwith thioflavin S (1% solution in water) for 8 min. Afterdifferentiation of the thioflavin S staining, sections werecounterstained with hematoxylin for 5 minutes. Two sets of pictures werecaptured simultaneously. A first set of pictures was captured underbrightfield illumination to obtain morphological details of the section;a second set of pictures was captured under a green, specific,fluorescent filter (fluorescein filter, Ex 495 nm, Em 525 nm). Imageanalysis to quantify the number of plaques and the area occupied bythese plaques was performed using Image Pro Plus software (MediaCybernetics, MD, USA).

The data from the histological ThioS studies is summarized in Table 13.Increases in the areas and numbers of plaques are indicated using “+”symbols, while decreases in the areas and numbers of the plaques areindicated using “−” symbols. Preliminary histochemical experiments usingThioS staining of brain sections indicated that both the number ofplaques and the area occupied by the plaques were decreased in micetreated with 30 mg/kg of compound BX.

Specifically, increases in the areas and numbers of plaques (relative tovehicle treated control) of 10 to 19.99% are scored as “+”. Decreases inthe areas and numbers of plaques of 10 to 19.99% are scored as “−”.Additionally, changes of 9.99% or less in either direction are scored as“0”.

TABLE 12 Effects of Compounds BX, BW and BZ on levels of Aβ in plasmaand brain Brain Dose Timepoint Plasma Abeta40 Abeta42 Compound (mg/kg)(weeks) Abeta40 Abeta42 soluble insoluble soluble insoluble BX 30  8wks + + −−− −−− −−− −− BX 100  8 wks ++ +++ + ++ + 0 BX 30 16 wks − − −−−−− 0 − BX 100 16 wks − 0 − −−− 0 −− BW 30  8 wks −−− −−− − −− −− 0 BW100  8 wks − − −− −−− −− −−− BW 30 16 wks −− −− + ++ − + BW 100 16 wks −− ++ + + ++ BZ 30  8 wks 0 0 0 −− 0 −−− BZ 100  8 wks ++ +++ ++ 0 0 − BZ30 16 wks 0 + 0 + + 0 BZ 100 16 wks ++ ++ −− 0 − +

TABLE 13 Histological effects of compounds BW and BX on numbers ofplaques and areas occupied by plaques ThioS Sites Analyzed SurfaceChange in Change in Com- Dose Timepoint Area Plaque Plaque pound (mg/kg)(weeks) (μm²) Number Area BX 30 16 wks 7,773,230 − − BX 100 16 wks7,803,230 0 + BW 30 16 wks 7,563,737 0 0 BW 100 16 wks 7,812,844 − 0

Example 5 Evaluation of Compounds Binding to NAC Peptide by MassSpectrometry

Recent findings have demonstrated that a high percentage of AlzheimerDisease (AD) patients also form Lewy bodies, most abundantly in theamygdala (Hamilton. 2000. Brain Pathol, 10:378; Mukaetova-Ladinska, etal. 2000. J Neuropathol Exp Neurol 59:408). Interestingly, the highlyhydrophobic non-amyloid component (NAC) region of α-synuclein has alsobeen described as the second most abundant component of amyloid plaquesin the brain of AD patients, after. Alpha-synuclein has been shown toform fibrils in vitro. Furthermore it binds to Aβ and promotes itsaggregation (Yoshimoto, et al. 1995. Proc Natl Acad Sci USA 92:9141). Itwas in fact originally identified as the precursor of the non-amyloidbeta (Aβ) component (NAD) of AD plaques (Ueda, et al. 1993. Proc NatlAcad Sci USA 90:11282; Iwai. 2000. Biochem Biophys Acta 1502:95;Masliah, et al. 1996. Am J Pathol 148:201). NAC is a 35 amino acid longpeptide with highly hydrophobic stretches which can self-aggregate andform fibrils in vitro. Moreover, these fibrils can efficiently seed theformation of Aβ fibrils in vitro (Han, et al. 1995. Chem Biol. 2:163-169; Iwai, et al. 1995. Biochemistry 34:10139). It is in factthrough its NAC domain that alpha-synuclein retains its fibrillogenicproperties. Modulating the properties of NAC or targeting NAC with thecompounds of the invention could therefore be a valid therapeutic avenueto inhibit the formation of protein aggregates and inclusions associatedwith alpha-synucleopathies, as well as the formation of aggregatesbetween the beta-amyloid peptide and NAC of alpha-synuclein.

The ability of the compounds of the present invention to bind to NACpeptide in aqueous solution was evaluated. The binding abilitycorrelates to the intensities of the peptide-compound complex peaksobserved by the Electrospray Mass Spectrum. Millipore distilleddeionized water was used to prepare all aqueous solutions. For pHdetermination a Beckman Φ36 pH meter fitted with a Corning Semi-MicroCombination pH Electrode was employed.

NAC (MW 3260.6 Da) at 20 μM was first analyzed at pH 7.40 and the usualsodium clusters was observed at +2, +3 and +4 at m/z 1335.5, 1116.7 and843.4 respectively. The optimal cone voltage was determined to be 20V.

Mass Spectrometry—

Mass spectrometric analysis was performed using a Waters ZQ 4000 massspectrometer equipped with a Waters 2795 sample manager. MassLynx 4.0(earlier by MassLynx 3.5) was used for data processing and analysis.Test compounds were mixed with disaggregated peptides in aqueous media(6.6% EtOH) at a 5:1 ratio (20 μM NAC: 100 μM of test compound or 40 μMNAC: 200 μM of test compound). The pH of the mixture was adjusted to 7.4(±0.2) using 0.1% NaOH (3-5 μL). Periodically, NAC peptide solution at20 μM or 40 μM was also prepared in the same fashion and run as control.The spectra were obtained by introducing the solutions to theelectrospray source by direct infusion using a syringe pump at a flowrate of 25 μL/min, and scanning from 100 to 2100 Da in the positivemode. The scan time was 0.9 sec per scan with an inter-scan delay of 0.1sec and the run time was 5 min for each sample. All the mass spectrawere sum of 300 scans. The desolvation and source temperature was 70° C.and the cone and capillary voltage were maintained at 20 V and 3.2 kVrespectively.

The total area under the peaks for the bound NAC-compound complexdivided by total area under the peaks for unbound NAC was determined foreach compound tested. The results are summarized in Table 14 below.

TABLE 14 NAC Peptide Binding Data Structure Binding Strength*  NaO₃SCH₂(CH₂)₂CH₂SO₃Na − NaO₃SOCH₂CH₂CH₂OSO₃Na − NH₂CH₂CH₂OSO₃H −H₂NCH₂CH₂CH₂OSO₃Na ++ H₂NCH₂CH₂SO₃H +

++

++

+

+++

*+++ = Strong; when the total binding is 120% and higher ++ = Moderate;when the total binding is between 120% and 70% + = Weak; when the totalbinding is between 70% and 30% − = None; when the total binding isbetween 30% and 0%

Example 6 Maintenance of Neuroblastoma SH-SY5Y, Treatments and HoechstStaining

A representative subset of compounds shown hereinbefore in Tables 2 and3 were tested for neuroprotective activity. Briefly, SH-SY5Y cells werecultured according to American Type Culture Collection (ATCC)recommendations. Briefly, cells were grown in a culture mediumcontaining 10% fetal bovine serum (Gibco), 1× non-essential amino acidsin a 1:1 mixture of Eagle's minimum essential medium (Sigma) and Ham'sF12 medium (Gibco).

Synthetic Aβ₄₂ (American Peptide) was resuspended in1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP), sonicated, and stored at −80°C. Preparations were thawed, dried under nitrogen gas, and dissolved in0.04 M Tris, 0.3 M NaCl (pH 7.4) at 120 μM. SH-SY5Y cells were seeded onglass coverslips in a 24-well plate at a density of 3×10⁵ cells/well.Treatments were performed the next day. Cells were incubated for 24hours with 12 μM Aβ₁₋₄₂, diluted in the culture medium from the 120 μMstock in the presence or absence of 240 μM NRM compounds (1:20 Aβ:drugratio).

Cell death was assessed using the DNA-binding dye Hoechst (33342)(Molecular Probes) to detect condensed or fragmented chromatin. Thecoverslips containing SH-SY5Y cells were stained with Hoechst 33342 (2μg/ml) for 10 min, fixed in 4% paraformaldehyde (Electron MicroscopyScience) for 30 minutes at room temperature, washed in PhosphateBuffered Saline (Gibco) and mounted onto glass slides using prolonganti-fade reagent (Molecular Probes). The nuclei were visualized using afluorescence microscope at 200× magnification. Live cells and cellsconsidered morphologically apoptotic were counted. Apoptotic nucleiappear condensed and occasionally fragmented. Five random fields werecaptured for each condition in a blinded fashion. Apoptotic and normalnuclei in each field were quantified by manual examination. The resultsare summarized in Table 15 below.

TABLE 15 Neuroprotective activity of selected compounds according to theinvention Inhibition of Aβ-induced toxicity (%) Compound ID <20% NM; NP;OQ; PJ; QD; QJ; QQ; N5; N9; N10; N11; N33; N47; N48; N49; N50; N53; N54;N55; N57; N58; N60; N61; N62; N63; N64; N65; N66; N67; N68; N70; N71;N72; N73; N74; N77; N79; N84; 20%-40% N14; N16; N37; N43; N44; N52; N75;N78; N80; N81; N83; N85; N86; N87; >40% N56; N59; N69; N82;

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims.

1. A compound of formula VIII, XII, XIV or XV:

wherein: R¹ is hydrogen, a substituted or unsubstituted cycloalkyl,heterocyclic, aryl, arylcycloalkyl, bicyclic or tricyclic ring, abicyclic or tricyclic fused ring group, or a substituted orunsubstituted C₂-C₁₀ alkyl group; R² is selected from a group consistingof hydrogen, alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl,arylalkyl, thiazolyl, triazolyl, imidazolyl, benzothiazolyl, andbenzoimidazolyl; R^(h) is hydrogen, benzyl, aryl-alkyl, aryl, or alkyl;R^(i) is hydrogen, substituted or unsubstituted aryl, substituted orunsubstituted benzyl, alkenyl, carbocyclic, heterocyclic, absent ortogether with R^(k), R^(k) or R^(m) may be linked to form a ringstructure; R^(j), R^(l), R^(m), R^(n), and R^(o) are each independentlyhydrogen, substituted or unsubstituted aryl, substituted orunsubstituted benzyl, alkyl, alkenyl, carbocyclic, heterocyclic, absentor together may be linked to form a ring structure; R^(s) is hydrogen orwhen n² is 3, R^(s) is (CH₂)₃—SO₃ ⁻X⁺; R^(q) and R^(r) are each selectedindependently from hydrogen or alkyl, R^(t) is hydrogen, alkyl, or aryl;R^(u) and R^(v) are each independently for each occurrence selected fromhydrogen, aryl, benzyl, alkyl, alkenyl, carbocyclic, heterocyclic, ortwo R^(u) or R^(v) groups on adjacent carbon atoms may form a doublebound, or together with the carbon atoms they are attached to forming acarbocyclic or heterocyclic ring; Y is SO₃ ⁻X⁺, OSO₃ ⁻X⁺, or SSO₃ ⁻X⁺;X⁺ is hydrogen, a cationic group, or an ester-forming group; L¹ is asubstituted or unsubstituted C₁-C₅ alkyl group or absent, B is C₁-C₅alkyl, alkenyl, or alkynyl group, and optionally fused with W when M isabsent; M is a covalent bond, amino, C₁-C₆ alkyl, alkenyl, alkynyl,carboxyl, oxy, amide, ester, thioether, thioester or absent; W is asubstituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl,arylalkyl, bicyclic or tricyclic ring, a bicyclic or tricyclic fusedring group, heterocyclic, thiazolyl, triazolyl, imidazolyl,benzothiazolyl, or benzoimidazolyl; v is 1, 2, 3, 4, 5, or 6; n² is 0,1, 2, or 3, selected such that three carbons are between the SO₃ ⁻X⁺group and the nitrogen atom in the ring; n³ is 4, 5, 6, or 7; and t¹ andt² are each single or double bonds, provided that both t¹ and t² are notboth double bonds: or a pharmaceutically acceptable salt, ester orprodrug thereof, provided that when Y is methyl, R¹ and R² are hydrogen,Y is SO₃ ⁻X⁺, and M is a covalent bond, B is not CH₂—CH(M-W)—CH₂. 2-3.(canceled)
 4. The compound of claim 1, wherein v is
 1. 5. The compoundof claim 1, wherein M is a covalent bond or a C₁-C₃ alkyl. 6-12.(canceled)
 13. The compound of claim 1, wherein B is —CH(M-W)—CH₂—CH₂—.—CH₂—CH(M-W)—CH₂—, or —(CH₂)—CH₂—CH(M-W)—. 14-16. (canceled)
 17. Acompound of Formula IX, X, XI, or XIII:

wherein: R¹ is a substituted or unsubstituted cycloalkyl, heterocyclic,aryl, arylcycloalkyl, bicyclic or tricyclic ring, a bicyclic ortricyclic fused ring group, or a substituted or unsubstituted C₂-C₁₀alkyl group; R² is selected from the group consisting of hydrogen,alkyl, mercaptoalkyl, alkenyl, alkynyl, cycloalkyl, aryl, arylalkyl,thiazolyl, triazolyl, imidazolyl, benzothiazolyl, and benzoimidazolyl;R³ is hydrogen or a protecting group; R^(a) is hydrogen, substituted orunsubstituted alkyl, aryl, heteroaryl, carboxyl, alkyloxycarbonyl, oraminocarbonyl; R^(b) and R^(c) are each selected independently fromhydrogen, substituted or unsubstituted alkyl, alkenyl, alkynyl,cycloalkyl, CONH₂, or R^(b), R^(c) and the carbon atom they are attachedto can form a substituted or unsubstituted cyclic structure of 4 to8-membered ring or a fused ring system; R^(d) is H or alkyl; R^(e) andR^(f) are each independently hydrogen, C₁-C₆ alkyl, or R^(e) and R^(f)taken together with the carbon they are attached to form a 3 to6-membered ring; R^(g) is independently selected for each occurrencefrom the group consisting of: hydrogen, alkyl, alkoxy, halogen, NO₂, andalkyl-SO₂; R^(p) and aa are each a natural or unnatural amino acidresidue; Ar is aryl or heteroaryl; P is a covalent bond, alkyl,alkyloxy, amino, alkylamino, sulfur, or alkylthio; Y is SO₃ ⁻X⁺, OSO₃⁻X⁺, or SSO₃ ⁻X⁺; X⁺ is hydrogen, a cationic group, or an ester-forminggroup; and Z is —(CH₂)₀₋₃—, —(CHOH)—, (CH₂)₁₋₃O(CH₂)₁₋₃, or a carbonylgroup; each of L¹ and L² is independently a substituted or unsubstitutedC₁-C₅ alkyl group or absent; L³ is a covalent bond, amino, C₁-C₆ alkyl,alkenyl, alkynyl, carboxyl, amide, aminoalkyl, ether, ester, thioether,thioester or absent; q is 1, 2, 3, 4, or 5; and n¹ is 0, 1, 2, or 3; ora pharmaceutically acceptable salt, ester or prodrug thereof. 18-61.(canceled)
 62. A compound of Table 3A or Table 3B, or a pharmaceuticallyacceptable salt, ester, or prodrug thereof.
 63. (canceled)
 64. A methodof treating or preventing an amyloid-related disease in a subjectcomprising administering to a subject in need thereof a compound ofclaim 1, or a pharmaceutically acceptable salt thereof, in an amounteffective to treat or prevent an amyloid related disease.
 65. The methodaccording to claim 64, wherein said amyloid-related disease isAlzheimer's disease, cerebral amyloid angiopathy, inclusion bodymyositis, macular degeneration, MCI, or Down's syndrome.
 66. (canceled)67. The method according to claim 64, wherein said amyloid-relateddisease is diabetes, AA amyloidosis, AL amyloidosis, ATTR-relatedamyloidosis, or hemodialysis related amyloidosis (β₂M). 68-70.(canceled)
 71. A method for treating or preventing an amyloid-relateddisease in a subject in need thereof, comprising administering to saidsubject a therapeutic compound of claim 17, or a pharmaceuticallyacceptable salt thereof, in an amount effective to treat or prevent anamyloid related disease.
 72. A method for treating or preventing anamyloid-related disease in a subject in need thereof, comprisingadministering to said subject a therapeutic compound of claim 62, orpharmaceutically acceptable salts thereof, in an amount effective totreat or prevent an amyloid related disease. 73-119. (canceled)
 120. Themethod according to claim 64, wherein said amyloid-related disease isfamilial amyloid polyneuropathy (FAP), senile systemic amyloidosis,Tenosynovium, familial amyloidosis, Ostertag-type, non-neuropathicamyloidosis, cranial neuropathy, hereditary cerebral hemorrhage,familial dementia, chronic dialysis, familial Creutzfeldt-Jakob disease,Gerstmann-Strä ussler-Scheinker syndrome, hereditary spongiformencephalopathies, prion diseases, familial Mediterranean fever,Muckle-Well's syndrome, nephropathy, deafness, urticaria, limb pain,cardiomyopathy, cutaneous deposits, multiple myeloma, benign monoclonalgammopathy, maccoglobulinaemia, myeloma associated amyloidosis,medullary carcinomas of the thyroid, isolated atrial amyloid, ordiabetes. 121-202. (canceled)
 203. The method according to claim 64,wherein the amyloid protein is amyloid λ, amyloid κ, amyloid κIV,amyloid γ, or amyloid γ1. 204-219. (canceled)
 220. The method accordingto claim 71, wherein said amyloid-related disease is Alzheimer'sdisease, cerebral amyloid angiopathy, inclusion body myositis, maculardegeneration, MCI, or Down's syndrome.
 221. The method according toclaim 71, wherein said amyloid-related disease is diabetes, AAamyloidosis, AL amyloidosis, ATTR-related amyloidosis, or hemodialysisrelated amyloidosis (β₂M).
 222. The method according to claim 71,wherein said amyloid-related disease is familial amyloid polyneuropathy(FAP), senile systemic amyloidosis, Tenosynovium, familial amyloidosis,Ostertag-type, non-neuropathic amyloidosis, cranial neuropathy,hereditary cerebral hemorrhage, familial dementia, chronic dialysis,familial Creutzfeldt-Jakob disease, Gerstmann-Strä ussler-Scheinkersyndrome, hereditary spongiform encephalopathies, prion diseases,familial Mediterranean fever, Muckle-Well's syndrome, nephropathy,deafness, urticaria, limb pain, cardiomyopathy, cutaneous deposits,multiple myeloma, benign monoclonal gammopathy, maccoglobulinaemia,myeloma associated amyloidosis, medullary carcinomas of the thyroid,isolated atrial amyloid, or diabetes.
 223. The method according to claim71, wherein the amyloid protein is amyloid λ, amyloid κ, amyloid κIV,amyloid γ, or amyloid γ1.
 224. The method according to claim 72, whereinsaid amyloid-related disease is Alzheimer's disease, cerebral amyloidangiopathy, inclusion body myositis, macular degeneration, MCI, orDown's syndrome.
 225. The method according to claim 72, wherein saidamyloid-related disease is diabetes, AA amyloidosis, AL amyloidosis,ATTR-related amyloidosis, or hemodialysis related amyloidosis (β₂M).226. The method according to claim 72, wherein said amyloid-relateddisease is familial amyloid polyneuropathy (FAP), senile systemicamyloidosis, Tenosynovium, familial amyloidosis, Ostertag-type,non-neuropathic amyloidosis, cranial neuropathy, hereditary cerebralhemorrhage, familial dementia, chronic dialysis, familialCreutzfeldt-Jakob disease, Gerstmann-Strä ussler-Scheinker syndrome,hereditary spongiform encephalopathies, prion diseases, familialMediterranean fever, Muckle-Well's syndrome, nephropathy, deafness,urticaria, limb pain, cardiomyopathy, cutaneous deposits, multiplemyeloma, benign monoclonal gammopathy, maccoglobulinaemia, myelomaassociated amyloidosis, medullary carcinomas of the thyroid, isolatedatrial amyloid, or diabetes.
 227. The method according to claim 72,wherein the amyloid protein is amyloid λ, amyloid κ, amyloid κIV,amyloid γ, or amyloid γ1.